The DOI matches its Crossref record; 129 of the 131 references it cites were found in a registry or an index, and 2 are not checked yet; 44 of the 86 sources its claims cite were checked for support.
Modeling Physiological Events in 2D vs. 3D Cell Culture
Open accessAbstract
Cell culture has become an indispensable tool to help uncover fundamental biophysical and biomolecular mechanisms by which cells assemble into tissues and organs, how these tissues function, and how that function becomes disrupted in disease. Cell culture is now widely used in biomedical research, tissue engineering, regenerative medicine, and industrial practices. Although flat, two-dimensional (2D) cell culture has predominated, recent research has shifted toward culture using three-dimensional (3D) structures, and more realistic biochemical and biomechanical microenvironments. Nevertheless, in 3D cell culture, many challenges remain, including the tissue-tissue interface, the mechanical microenvironment, and the spatiotemporal distributions of oxygen, nutrients, and metabolic wastes. Here, we review 2D and 3D cell culture methods, discuss advantages and limitations of these techniques in modeling physiologically and pathologically relevant processes, and suggest directions for future research.
Verified 2026-10-07 · 2 references not checked yet
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Claims the paper makes
19 No full text of the sources · 16 Partly supported · 12 Own interpretation · 10 Supported · 7 Couldn’t confirm · 3 Supported so far · 1 Not supported · accuracy not yet measured
Every source the paper cites for this claim was checked and supports it.
In the paper
“These behaviors include cell differentiation, migration, growth, and mechanics, all of which are impacted by their biochemical and biomechanical microenvironment (57).”
§ 2D vs. 3D Cell Culture > Abstract
What the sources say
- Supportedcites [57] source
In “Microengineering Meets Cell Biology” (page not provided), the paper says the mechanical environment is critical for cell growth and tissue differentiation, and describes chemical gradients used to study cell motility. The same section describes how ECM patterning and substrate conditions affect cell shape and tissue contraction, supporting the claim about mechanics. (LLM, conf=0.90)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“For instance, some important characteristics of cancer cells cannot be appropriately modeled in 2D cultures (22).”
§ 2D vs. 3D Cell Culture > Abstract
What the sources say
- Could not verifycites [22] sourceExperimental — accuracy not yet measured
The Experimental Section describes culturing NIH-3T3 fibroblasts in porous beads, and the paper’s discussion of three-dimensional matrices concerns cell viability. Searches for cancer cells and comparisons with 2D cultures found no passage addressing whether 2D cultures inadequately model cancer-cell characteristics; page numbers were not available in our copy. (LLM, conf=0.74)
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Could not verify
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Results from 3D studies demonstrate that increasing the dimensionality of extracellular matrix (ECM) around cells from 2D to 3D can significantly impact cell proliferation, differentiation, mechano-responses, and cell survival (2,10,41).”
§ 2D vs. 3D Cell Culture > Abstract
What the sources say
- Not checked — no full text of the sourcecites [2] source
- Partly supportedcites [10] sourceExperimental — accuracy not yet measured
The Introduction (page not provided) reports that 3D matrices can alter cell phenotype and metabolism and discusses proliferation, differentiation, and resistance to toxic effects. It does not address mechano-responses. The Discussion (page not provided) also states that this study found no evidence that moving cells from 2D plastic to 3D collagen affected their health or viability, so the paper does not support the full list as a finding of this study. (LLM, conf=0.84)
- Partly supportedcites [41] sourceExperimental — accuracy not yet measured
The Results (§3.3; page unavailable) show that cells remained viable and proliferated on the 3D GelMA scaffolds. The Introduction (§1) discusses scaffold cues affecting differentiation, but the paper does not establish that changing the ECM from 2D to 3D significantly changes all four listed outcomes; its mechanical measurements concern the scaffolds, not cellular mechano-responses. (LLM, conf=0.84)
boosteddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“The sandwich culture method, e.g., placing cells between two layers comprised of ECM, polyacrylamide, or collagen, has long been demonstrated to produce hepatocyte cell cultures, where morphology and function more accurately reproduce in vivo behavior (9,29,34,65,73).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Not checked — no full text of the sourcecites [9] source
- Partly supportedcites [29] sourceExperimental — accuracy not yet measured
The opening summary (p. 1043) reports that rat hepatocytes between two collagen layers maintained albumin mRNA at levels similar to normal liver, supporting the claim about preserved function. The paper does not demonstrate that sandwich cultures reproduce in vivo morphology more accurately, and it tests collagen rather than the other materials listed in the citing sentence. (LLM, conf=0.85)
- Not checked — no full text of the sourcecites [34] source
boosteddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Cells in our body perform bioactivities in response to the stimulation from a highly complex, 3D microenvironment (15,98,112).”
§ 2D vs. 3D Cell Culture > Considerations
What the sources say
- Supportedcites [15] source
The Biomimetic Approach section says that cells in living organisms inhabit a biologically complex microenvironment, respond to its molecular and physical cues, and remodel it. It contrasts this native context with standard 2D culture and describes the relevant in vivo environment as dynamic and 3D. (LLM, conf=0.94)
- Partly supportedcites [98] sourceExperimental — accuracy not yet measured
The Results (Cell interactions with scaffolds; page unavailable) show that endothelial-cell infiltration responds to porosity, RGD peptide, and S1P delivery in three-dimensional engineered scaffolds. This supports the general idea that a cell’s surroundings influence its activity, but the paper does not establish the broader claim about cells throughout the body responding to a highly complex natural 3D microenvironment. (LLM, conf=0.84)
- Not checked — no full text of the sourcecites [112] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“The introduction of 3D cell culture approaches aimed at modeling the in vivo interactions of tissues and organs has opened new possibilities in studying the underlying biochemical and biomechanical signals (46,57).”
§ 2D vs. 3D Cell Culture > Considerations
What the sources say
- Supportedcites [46] source
The “Biomaterials to create 3D tumor models” section says 3D cultures better resemble tumors in vivo and help researchers understand cancer biology. Its subsections describe using 3D models to study cellular interactions, soluble microenvironmental cues, and matrix mechanics; “Future directions and considerations” also discusses organ-on-chip models. The paper supports the claim in the context of cancer models. (LLM, conf=0.87)
- Supportedcites [57] source
The “3D Cell Culture” section describes a new wave of 3D models designed to mimic organ structure, mechanical properties, and biochemical functions. In “From 3D Culture to Organs-on-Chips,” the paper says these microengineered approaches opened new possibilities for modeling organ-level structures and incorporating mechanical and chemical signals. The claim is supported as a description of these advanced 3D approaches, rather than conventional gel cultures. (LLM, conf=0.88)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“This device was able to maintain spheroid cultures for up to 2 wk (110).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Partly supportedcites [110] sourceExperimental — accuracy not yet measured
The Results and discussion section, under “Long-term culture of spheroids in hanging drops” (page unavailable), reports viable spheroids after 12 days of culture. This supports long-term maintenance with the device, but the copy does not establish the claimed two-week (14-day) duration. (LLM, conf=0.85)
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Partly supported
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“Hepatocytes in the liver are surrounded by a complex ECM, and several researchers have shown that conventional 2D culture methods, flasks with or without ECM protein coatings, do not establish well enough the complex microenvironment that hepatocytes need to function (29,34,73).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Supportedcites [29] source
The Introduction (p. 1043) says hepatocytes rapidly degenerate under classical culture conditions and that adding complex extracellular matrix can prolong their function. The Results (p. 1046) show that albumin mRNA declined rapidly on a single collagen-gel layer but was maintained in collagen-sandwich culture, supporting the claim that conventional flat cultures inadequately sustain hepatocyte function. (LLM, conf=0.88)
- Not checked — no full text of the sourcecites [34] source
- Not checked — no full text of the sourcecites [73] source
boosteddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“This induced polarity may alter the functions of native cells with regard to spreading, migrating, and sensing soluble factors and other microenvironmental cues (61).”
§ 2D vs. 3D Cell Culture > Considerations
What the sources say
- Could not verifycites [61] sourceExperimental — accuracy not yet measured
The ‘Engineered microenvironments regulate LA/C-C epitope accessibility’ section reports that substrate properties affect cell spreading and nuclear-lamina polarization. The ‘Physiological implications’ section links that polarization to engineered niches, but neither section says that induced polarity in pseudo-3D models alters native-cell spreading, migration, or sensing of soluble factors and other cues. Searches for pseudo-3D models, migration, and soluble-factor sensing did not locate such a passage. (LLM, conf=0.73)
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Could not verify
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“To control cell shape in 2D cell culture, micro-patterned substrates, such as cell-adhesive islands (30), microwells (121), and micropillars (40), have been created to customize the 2D shape of cells and help study the effects of cell shape on bioactivities.”
§ 2D vs. 3D Cell Culture > Considerations
What the sources say
- Not checked — no full text of the sourcecites [30] source
- Not checked — no full text of the sourcecites [121] source
- Supportedcites [40] source
Citation (40) is attached to the micropillars example. The paper reports fabricating micropost arrays and using them to study changes in cell morphology and stem-cell differentiation (Abstract; Online Methods, ‘Design and fabrication of PDMS micropost arrays’; page unavailable). (LLM, conf=0.88)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“The efficiency of the surface attachment was also shown to be higher on rounded surfaces compared with smooth surfaces.”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Could not verifycites [113] sourceExperimental — accuracy not yet measured
The Materials and Methods passage describes breast-cell cultures on plastic, collagen-coated dishes, and in basement membrane, but does not compare attachment to rounded and smooth surfaces. A search for surface attachment efficiency and that shape comparison found no passage addressing the claim. Page numbers were not available in our copy. (LLM, conf=0.76)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Could not verify
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“The effect of cell polarization in 2D cell culture can be mitigated using a sandwich culture method that adds a layer of ECM atop the cells (9,29,34,65,73) to eliminate apical-basal polarity and provide a mimic of 3D ECM.”
§ 2D vs. 3D Cell Culture > Considerations
What the sources say
- Not checked — no full text of the sourcecites [9] source
- Partly supportedcites [29] sourceExperimental — accuracy not yet measured
The Hepatocyte Culture methods describe adding a second collagen-gel layer over the cells, supporting the sandwich-culture method. But the discussion proposes that this configuration establishes proper cellular polarity, with two basolateral surfaces separated by an apical belt—not that it eliminates apical-basal polarity. The passages do not explicitly characterize it as a mimic of 3D ECM; page numbers were not supplied. (LLM, conf=0.87)
- Not checked — no full text of the sourcecites [34] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Engineered 2D surface patterning and modification creates a 2D microenvironment with unique biochemical factors, topography, stiffness, and mechanical load for cell culture (72,81).”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Not checked — no full text of the sourcecites [72] source
- Partly supportedcites [81] sourceExperimental — accuracy not yet measured
The Results (§3.1; page unavailable) describe cells cultured on 2D hydrogel surfaces with an adhesive peptide and a specified modulus. The Discussion (§4; page unavailable) says the system presents biochemical and mechanical matrix cues. The paper does not describe engineered surface patterning or topography, or an applied mechanical load, as features of its 2D culture system. (LLM, conf=0.86)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Every source the paper cites for this claim was checked and supports it.
In the paper
“GEL and unattached EB cultures produced similar cluster morphology, with defined boundaries and the occasional cavities.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Supportedcites [91] source
The Results section (‘Cell Growth in Differentiation Systems’; page not specified) says GEL cell organization was similar to standard EB culture, with bodies that commonly had defined boundaries and occasionally contained cavities. (LLM, conf=0.94)
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“However, the ESCs cultured on glass slides (2D) coated with collagen type I had a faster rate of differentiation than in 3D cultures (EBs and GELs).”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
What the sources say
- Supportedcites [91] source
The Materials and Methods section identifies the 2D system as mouse ESCs cultured on collagen type I-coated glass slides and the 3D systems as gels and embryoid bodies. The Discussion states that 2D adherent cultures induced overall differentiation more quickly than the 3D configurations. (LLM, conf=0.95)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“B: the system could recreate organ-level functions, such as an inflammatory response bacteria.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
What the sources say
- Supportedcites [33] source
The “Microphysiological organ models” section says the lung-on-a-chip reconstituted organ-level responses, including immune-cell recruitment and phagocytic activity in response to bacteria. The supplied passage does not give a page number. (LLM, conf=0.93)
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“A key difference in the morphology of GEL cultures was the presence of elongated masses along the edges of the gel form.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Supportedcites [91] source
The Results section (Cell Growth in Differentiation Systems) describes elongated bodies along the edges of the gel construct as unique to GEL cultures. This directly supports the cited morphological difference. (LLM, conf=0.94)
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“By day 12, unattached EB cultures had higher expression levels of the intermediate filaments lamin, keratin 8, and vimentin (91).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Supportedcites [91] source
The Results section (“Matrix Remodeling”; page unavailable in our copy) identifies lamin (LMNA), keratin 8 (KRT8), and vimentin (VIM) as intermediate filaments and reports higher expression in EB cultures at day 12. This supports the cited comparison. (LLM, conf=0.92)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“These 3D scaffolds and associated cell-encapsulation techniques provide the valuable tools for helping us decipher how the ECM affects the fate of cells (5, 13, 14, 25-27, 68, 77, 80, 87, 88, 92, 108).”
§ 2D vs. 3D Cell Culture > Biopolymer scaffolds.
What the sources say
- Supportedcites [5] source
The Results and Discussion describes 3D hydrogel encapsulation as a way to test how chemical features of the cell environment affect hMSC differentiation while holding cell density and morphology consistent. It also calls the approach a methodology for improving understanding of how extracellular interactions affect cell activities. Page numbers are not supplied in our copy. (LLM, conf=0.93)
- Not checked — no full text of the sourcecites [13] source
- Partly supportedcites [14] sourceExperimental — accuracy not yet measured
The Materials and Methods describe cell encapsulation in 3D hydrogel networks, and the Results and Discussion report how network properties relate to cell viability and neocartilage formation. This supports the scaffolds-and-encapsulation part of the claim, but the paper does not directly examine how the extracellular matrix affects cell fate; it chiefly examines how the networks affect cells and matrix distribution. Page numbers were not available in our copy. (LLM, conf=0.82)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
The sources with a verdict support this claim; the rest weren’t checked yet, have no full text we could read, or their verdicts aren’t rated yet.
In the paper
“This approach has been developed in the research of cancer (7, 76, 102), stem cells (17, 120), and many organs (such as liver, kidney, heart, etc.) (3,16,64,75).”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
What the sources say
- Supportedcites [7] source
The citation to this paper is among those attached to cancer research, not those attached to stem cells or organs. The Introduction describes a spheroid-based method for studying cancer cell invasion, and the Short Abstract says the method evaluates cancer cell invasion from spheroids into a 3D matrix. (LLM, conf=0.93)
- Supportedcites [76] source
The Introduction describes spheroid culture in cancer research, and the Conclusions report a technology for generating cancer cell spheroids. This source is cited in the cancer-reference group; separate citations accompany the stem-cell and organ examples. (LLM, conf=0.92)
- Could not verifycites [102] sourceExperimental — accuracy not yet measured
Section 6.1 describes a miniature liverlike construct made from collagen rods, and Section 6.3 describes stem cells cultured within spherical hydrogel microgels. Neither establishes that spheroid cell culture was developed for research on liver, kidney, heart, or the other named subjects. Searches for “spheroid,” “kidney,” and “heart” found no passage establishing that claim; page numbers were unavailable. (LLM, conf=0.73)
- Supportedcites [17] source
The cited paper directly supports the stem-cell portion of the sentence: its Introduction says spheroids have been used in stem cell biology, and Section 2.4 describes spheroid cultures of pluripotent and neural stem cells. The sentence groups separate citations with the cancer and organ examples; those examples need not be supported by this stem-cell source. (LLM, conf=0.91)
- Not checked — no full text of the sourcecites [120] source
- Not checked — no full text of the sourcecites [3] source
- Not checked — no full text of the sourcecites [16] source
- Not checked — no full text of the sourcecites [64] source
- Not checked — no full text of the sourcecites [75] source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“C: the system was also used as a model for lung diseases like pulmonary edema.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
What the sources say
- Supportedcites [33] source
The Microphysiological organ models section says the lung-on-a-chip system was engineered to model the development and progression of pulmonary oedema. The Toxicity screening section specifies that it simulated pulmonary oedema caused by IL-2 drug toxicity. (LLM, conf=0.94)
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Free EBs had more dynamic changes in the genotypic expression levels of cytoskeletal genes over the same 12-day span compared with the GEL cultures.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Partly supportedcites [91] sourceExperimental — accuracy not yet measured
The Results section (Matrix Remodeling; page unavailable) reports that intermediate-filament gene expression rose in both EB and GEL cultures from day 4 to days 8 and 12, with higher levels in EBs at day 12. It does not establish more dynamic changes across cytoskeletal genes generally: the measured microtubule gene was unaffected by time or culture condition. (LLM, conf=0.82)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Partly supported
In the paper
“The patterned PLLA surfaces were shown to have a faster rate of differentiation than non-patterned PLLA.”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Not checked — no full text of the sourcecites [18] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“Last but not least, cells cultured on the stiffest matrix (E ϭ 25-40 kPa) expressed fourfold higher osteogenic factor messages.”
§ 2D vs. 3D Cell Culture > Altering substrate stiffness.
What the sources say
- Not checked — no full text of the sourcecites [32] source
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“In response to variations in microtopography, cells tend to exhibit an increase in filopodia and microspikes, which play important roles in cell migration and neurite outgrowth (106).”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Not checked — no full text of the sourcecites [106] source
plainconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“Cultures on the softest gels with Young's modulus (E) similar to the brain (0.1-1 kPa) exhibited the highest expression of neuron-specific markers.”
§ 2D vs. 3D Cell Culture > Altering substrate stiffness.
What the sources say
- Not checked — no full text of the sourcecites [32] source
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“Cells on a substrate with a moderate stiffness similar to muscle tissue (E ϭ 8 -17 kPa) expressed sixfold higher levels of myogenic factor messages.”
§ 2D vs. 3D Cell Culture > Altering substrate stiffness.
What the sources say
- Not checked — no full text of the sourcecites [32] source
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“The sandwich method was shown to reduce the diffusion of oxygen sufficiently so that an 80% survival rate was found over 5 days compared with a 32% survival rate over 2 days in a traditional mixed-cell population culture (12).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Not checked — no full text of the sourcecites [12] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“As for lipid production, it was found that, at later time points, patterned PLLA surfaces produced the highest amount of lipids, followed by PS, and then non-patterned PLLA.”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Not checked — no full text of the sourcecites [18] source
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“However, cells cultured on patterned and non-patterned PLLA surfaces showed a lower rate of differentiation compared with cells on tissue culture-treated polystyrene (PS), a control for the experiment.”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Not checked — no full text of the sourcecites [18] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“The microenvironment of cells is not only affected by microtopography but also by the stiffness of the substrate that cells adhere to, which affects bioactivities ranging from migration to differentiation (28).”
§ 2D vs. 3D Cell Culture > Altering substrate stiffness.
What the sources say
- Not checked — no full text of the sourcecites [28] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“In a typical 2D cell culture, cells are exposed to a homogenous surface devoid of any defects that could influence their development (109).”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Not checked — no full text of the sourcecites [109] source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“For example, Engler et al. (32) demonstrated that the differentiation of mesenchymal stem cells (MSCs), which possess the ability to develop into neurons, myoblasts, and osteoblasts, can be regulated by substrate stiffness.”
§ 2D vs. 3D Cell Culture > Altering substrate stiffness.
What the sources say
- Not checked — no full text of the sourcecites [32] source
boosteddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“They determined that low levels of oxygen (9%) are ideal for low density (Ͻ20,000 cells/cm 2 ) hippocampal neuron cultures, since higher levels become toxic.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Not checked — no full text of the sourcecites [12] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Cells in 2D and 3D microenvironments interact differently with their surroundings due to differences in the cues, mechanical and chemical, that they experience”
§ 2D vs. 3D Cell Culture > Mechanical Responses
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Cells in spheroids often initiate different characteristics than monolayer cells, such as the deposition of ECM, secretion of growth factors, and gene expression profiles.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: unclear — It is unclear why the paper says it holds.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“However, a standard system for 3D cultures will be difficult to implement since different cell types will most likely be better matched for certain 3D systems.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“They found that similar forces are generated even as the stiffness of the matrix is changed.”
§ 2D vs. 3D Cell Culture > Mechanical Responses
What the sources say
- Not checked — no full text of the sourcecites [105] source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: Accurate measurements of the forces exerted on the extracellular matrix are necessary for understanding physiological processes such as cancer or immune cell migration.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“These results indicate that, even with similar morphologies, the gene expression of each 3D culture is specific in adaptation to its microenvironment.”
§ 2D vs. 3D Cell Culture > Techniques
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“Steinwechs et al. (105) quantified, for the first time, the forces resulting from a breast carcinoma cell that migrates within a 3D collagen fibril matrix.”
§ 2D vs. 3D Cell Culture > Mechanical Responses
What the sources say
- Not checked — no full text of the sourcecites [105] source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: Accurate measurements of the forces exerted on the extracellular matrix are necessary for understanding physiological processes such as cancer or immune cell migration.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“The difficulty in growing physiologically similar 3D tissue or organ cultures stems from finding viable cell types, biocompatible scaffold materials, and a scaffold design that supports the development of an ECM and vascularization.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
In the paper
“This is in contrast to 2D matrices whereby physical forces increase as the matrix stiffness is increased (115).”
§ 2D vs. 3D Cell Culture > Mechanical Responses
What the sources say
- Not checked — no full text of the sourcecites [115] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- Rests on an assumption: Accurate measurements of the forces exerted on the extracellular matrix are necessary for understanding physiological processes such as cancer or immune cell migration.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In 3D cell culture, multicellular spheroids are closer to living tissues in vivo in terms of biostructural and biofunctional properties compared with conventional 2D planar cell cultures.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
- It may state two separate claims as one. 16
- Probably not a substantial claim. 14
At least one cited source doesn’t support this claim or points to a different paper, and none confirmed it — see the verdicts below.
In the paper
“PDMS wells with a triangular pyramid shape opening promoted the growth of spheroids, with some control over the size achievable.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Disputedcites [110] sourceExperimental — accuracy not yet measured
The Experimental section (‘Plate design, fabrication, and hanging drop formation’; page unavailable) says the plate is made of polystyrene, not PDMS, and describes circular access holes and plateaus by diameter, not triangular-pyramid openings. The Results section (‘Formation of hanging drops for spheroid culture’; page unavailable) does support spheroid formation, while the Introduction says the platform forms uniformly sized spheroids. The claimed well material and opening shape are contrary to the paper’s design description. (LLM, conf=0.90)
Checked by a reasoning model — may be wrong.
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Disputed
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“To overcome this limitation, novel 3D cell culture platforms are being created to better mimic in vivo conditions and are sometimes called spheroid or organoid culture, as described below (37,61,84,98,117,118).”
§ 2D vs. 3D Cell Culture > Abstract
What the sources say
- Not checked — no full text of the sourcecites [37] source
- Could not verifycites [61] sourceExperimental — accuracy not yet measured
The Methods section describes cells plated on hydrogels, microposts, and adhesive micro-islands, while ‘Engineered microenvironments regulate LA/C-C epitope accessibility’ examines their effects on the nuclear lamina. Searches for spheroid or organoid culture and for 3D platforms that mimic in vivo conditions found no passage addressing the claim; the paper’s engineered substrates alone do not establish it. (LLM, conf=0.77)
- Not checked — no full text of the sourcecites [84] source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“They demonstrated that cells preferred to round up on smooth-surfaced PLLA substrates and stretch on rough surfaces.”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Could not verifycites [113] sourceExperimental — accuracy not yet measured
The Materials and Methods section describes breast cells cultured in basement membrane, and the Abstract discusses integrin–EGFR signaling. Searches for PLLA, surface roughness, and rounded versus stretched cells found no passage addressing the claimed substrate comparison. (LLM, conf=0.75)
boostedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Could not verify
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Torisawa et al. (110) created a novel chip system utilizing microfluidics that allows spheroids to grow in a controlled environment for long-term culture.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Partly supportedcites [110] sourceExperimental — accuracy not yet measured
The Introduction and the Results and discussion section on long-term culture support that the authors developed a platform for stable, long-term spheroid culture. But the Experimental section describes their system as an injection-molded 384 hanging-drop plate, not a microfluidic chip; the Introduction discusses microfluidic devices as other approaches. Torisawa is a coauthor, although Tung is the paper’s first author. (LLM, conf=0.88)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Their results showed that the sandwich culture could maintain a proper albumin production rate for at least 6 wk, whereas the one-gel system already had rates that were tenfold lower than normal by the end of the first week (29).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Partly supportedcites [29] sourceExperimental — accuracy not yet measured
The Results (p. 1045) say sandwich cultures maintained albumin production for at least six weeks and that production rates differed roughly tenfold between the sandwich and single-gel systems by the end of week one. The paper does not say the single-gel production rate was tenfold lower than normal; its comparison with normal liver concerns albumin mRNA levels, not production rates. (LLM, conf=0.90)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Partly supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“Cell growth in 2D monolayers allows for access to a similar amount of nutrients and growth factors present in the medium, which results in homogenous growth and proliferation (31).”
§ 2D vs. 3D Cell Culture > Considerations
What the sources say
- Supportedcites [31] source
The ‘Characteristics of 3D Cell Cultures Versus the Traditional 2D Cell Culture’ section says all cells in a 2D monolayer receive a homogeneous amount of nutrients and growth factors and that the monolayer is mainly composed of proliferating cells. Its contrast with the varied cell states in 3D cultures supports the paraphrase about homogeneous growth and proliferation. (LLM, conf=0.87)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“In 2D cultures, there are decreases in transcriptional genes and rapid changes in morphology (29,34).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Partly supportedcites [29] sourceExperimental — accuracy not yet measured
The Results (p. 1047) show that albumin-gene transcription declined over seven days in hepatocytes cultured on a single collagen gel. The Discussion (p. 1050) mentions a change in actin-filament organization when hepatocytes are sandwiched between collagen gels, but does not establish rapid morphological changes in the single-gel culture. The claim also generalizes a finding about specific genes in rat hepatocytes to 2D cultures broadly. (LLM, conf=0.86)
- Not checked — no full text of the sourcecites [34] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Partly supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“The effect of microtopography on cell migration was further supported by Wan et al. (113), whose experiments also revealed that pseudopods extending from cells could encroach into pits in culture surfaces.The less rigid pseudopods could act as anchors for the cell and influence its migration patterns, thereby providing a contact guidance (113).”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Could not verifycites [113] sourceExperimental — accuracy not yet measured
The Abstract describes β1-integrin and EGFR signaling in three-dimensional breast-cell cultures, not migration over patterned surfaces. Searches for pseudopods, pits, microtopography, anchors, and contact guidance found no passage addressing the attributed experiments. The cited paper is by Wang et al., whereas the citing sentence names Wan et al. (LLM, conf=0.75)
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Could not verify
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Surfaces of the aggregated spheroids, for example, have the highest levels of proliferation, whereas the interior of the 3D cell bodies possesses the highest number of quiescent or necrotic cells (31).”
§ 2D vs. 3D Cell Culture > Considerations
What the sources say
- Partly supportedcites [31] sourceExperimental — accuracy not yet measured
In “Characteristics of 3D Cell Cultures Versus the Traditional 2D Cell Culture” (page unavailable), the paper says spheroid outer layers are mainly proliferating and core cells tend to be quiescent or hypoxic. It lists necrotic cells among the cells in spheroids, but does not say they are concentrated in the interior or establish where the highest numbers occur. (LLM, conf=0.86)
plaincorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: unclear — It is unclear whether the paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“They continued on to use these plates as a high throughput method for testing anticancer drug sensitivities on H9 human epithelial carcinoma cells.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Partly supportedcites [55] sourceExperimental — accuracy not yet measured
The Materials and Methods section (Hanging Drop Formation and Spheroid Culture; page unavailable) confirms that Hsiao et al. used 384 hanging-drop plates to culture cancer-containing spheroids. It describes PC-3Luc GFP prostate cancer cells in a co-culture, not H9 cells, and the experiment assessed long-term culture rather than anticancer drug sensitivity. (LLM, conf=0.87)
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Biopolymers from non-mammal tissues, such as alginate and chitosan, are also used for making 3D scaffolds (23,63,68,84).”
§ 2D vs. 3D Cell Culture > Biopolymer scaffolds.
What the sources say
- Partly supportedcites [23] sourceExperimental — accuracy not yet measured
The Introduction describes alginate as a naturally derived polysaccharide, and the Conclusion reports using alginate hydrogels for in vitro 3D cell culture. This supports alginate as a 3D scaffold material, but the paper does not identify its source as non-mammalian or address chitosan. (LLM, conf=0.86)
- Not checked — no full text of the sourcecites [63] source
- Not checked — no full text of the sourcecites [68] source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Partly supported
Every source the paper cites for this claim was checked and supports it.
In the paper
“To overcome this, one group, Hsiao et al. (55), designed a plate using micro-ring structures to stabilize the drops, allowing for 14day culture of prostate cancer cells.”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Supportedcites [55] source
The Introduction describes a modified hanging-drop plate with micro-ring features designed to stabilize droplets. Materials and Methods reports 14-day culture of spheroids containing PC-3Luc GFP prostate cancer cells, and Results and Discussion reports that the drops remained stable throughout that period. The cells were grown in co-culture, a detail the citing sentence omits but does not contradict. (LLM, conf=0.94)
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
- Supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Among the most commonly used natural polymers are hyaluronic acid, gelatin, collagen, and chondroitin sulfate (14, 108).”
§ 2D vs. 3D Cell Culture > Biopolymer scaffolds.
What the sources say
- Partly supportedcites [14] sourceExperimental — accuracy not yet measured
The Introduction (page unavailable) identifies hyaluronic acid and collagen as materials investigated for cartilage scaffolds, supporting their use as natural biomaterials. It does not identify gelatin as a commonly used material, and mentions chondroitin sulfate as a cartilage component rather than as a commonly used polymer. It also does not establish that the four named materials are among the most commonly used. (LLM, conf=0.84)
- Not checked — no full text of the sourcecites [108] source
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
- Partly supported
The cited sources support only part of this claim — read each source’s verdict below.
In the paper
“Pineda et al.(91) demonstrated that OCT4 in mouse ESCs decreased in both 2D and 3D cultures, showing a loss of pluripotency, whereas NES (ectoderm) and BRACHY-T (mesoderm) markers were shown to increase.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
What the sources say
- Partly supportedcites [91] sourceExperimental — accuracy not yet measured
In Results, ‘Pluripotency and Germ Lineage Differentiation’ (page unavailable), OCT4 markedly decreased after four days in the 2D slide and 3D gel and embryoid-body cultures, indicating loss of pluripotency. The same section says NESTIN (NES) and BRACHY-T varied over time and were highest at day 4, rather than showing a general increase; ‘Functional Gene Groupings’ also reports that NES remained very low. (LLM, conf=0.85)
boosteddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Partly supported
The cited sources that could be checked couldn’t confirm or dispute this claim.
In the paper
“Their results from 2D and 3D spheroid culture indicated spheroids were more resistant to one anti-cancer drug, 5-fluorouracil, which inhibits proliferation, whereas 2D cultures were more resistant to a different anticancer drug, tirapazamine, which is cytotoxic in hypoxic conditions (111).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Could not verifycites [111] sourceExperimental — accuracy not yet measured
The Introduction discusses stem-cell culture platforms and small-molecule screens, but does not report a 2D-versus-spheroid drug-response comparison. Searches for spheroids, 5-fluorouracil, and tirapazamine found no passage addressing the claimed resistance results; the indexed passages provide no page numbers. (LLM, conf=0.76)
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
- Could not verify
In the paper
“LeCluyse et al. (73) found that the polarity of hepatocytes was also affected when cultured on only single-layer gels.When a sandwich culture was used, a normal distribution of the apical membrane markers was found at the canalicular membrane, and bile canaliculi were also well established (73).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Not checked — no full text of the sourcecites [73] source
boosteddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
In the paper
“The results indicated that the micro-patterning and type of surface both affected the rate of differentiation of cells (18).”
§ 2D vs. 3D Cell Culture > Micro-pattering.
What the sources say
- Not checked — no full text of the sourcecites [18] source
hedgeddirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“Overall, 3D platforms are likely to provide an increasingly attractive alternative for 2D cell culture as the technology develops to enable a wider range of processes.”
§ 2D vs. 3D Cell Culture > Abstract
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: fails — It may be too vague to check against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may be too vague to check against evidence.
- Rests on an assumption: 2D cell culture approaches can still recapitulate in vivo behavior for many bioactivities.
In the paper
“Micropatterning can also be used to pattern areas or to create “microwells” for the controlled growth of spheroids (i.e., agarose) (35).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Not checked — no full text of the sourcecites [35] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“These methods allow for a more physiologically relevant environment to investigate cell and tissue behavior.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim.
- Rests on an assumption: Three-dimensional studies are becoming increasingly necessary to replicate the behaviors of some in vivo conditions.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“different cell types will most likely be better matched for certain 3D systems.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
hedgedcorrelational- Asserted: passes — The authors put it forward as true.
- Checkable: fails — It may be too vague to check against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: fails — The paper may give no reason why it holds.
Probably not a substantial claim. It may be too vague to check against evidence. The paper may give no reason why it holds.
In the paper
“Additionally, microwells made out of a non-adhesion promoting material, such as agarose, can be used to grow several arrays of spheroids at once (86).”
§ 2D vs. 3D Cell Culture > Techniques
What the sources say
- Not checked — no full text of the sourcecites [86] source
plaindirect- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: fails — The paper’s conclusions may not depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. The paper’s conclusions may not depend on it.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In some cases, 2D studies are more feasible, and the results can be meaningful.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
plain- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: fails — The paper may give no reason why it holds.
Probably not a substantial claim. The paper may give no reason why it holds. It may state two separate claims as one.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“In conclusion, the situation and questions at hand will determine whether a 2D or 3D culture method is best suited.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
plainconditional- Asserted: passes — The authors put it forward as true.
- Checkable: fails — It may be too vague to check against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may be too vague to check against evidence.
A conclusion the paper draws from its own results — it cites no other source for it, so there is none to check. The quote shows it is in the paper.
In the paper
“However, in other cases, 3D studies will yield more accurate results and can be done in combination with 2D studies.”
§ 2D vs. 3D Cell Culture > Conclusions and Discussion
hedgedconditional- Asserted: passes — The authors put it forward as true.
- Checkable: passes — It can be checked against evidence.
- Load-bearing: passes — The paper’s conclusions depend on it.
- Based: passes — The paper gives a reason why it holds.
Probably not a substantial claim. It may state two separate claims as one.
The paper relies on this without support.
Under some circumstances, 2D systems produce cell bioactivities that deviate appreciably from the in vivo response.
“Although these approaches are well-accepted and have significantly advanced our understanding of cell behavior, growing evidence now shows that, under some circumstances, the 2D systems can result in cell bioactivities that deviate appreciably the in vivo response.”
The paper relies on this without support.
2D cell culture approaches can still recapitulate in vivo behavior for many bioactivities.
“additionally, 2D cell culture approaches can still recapitulate in vivo behavior for many bioactivities, while new advances in substrate”
The paper relies on this without support.
A universal 3D platform does not currently exist.
“the platform of choice is often dictated by the specific process of interest, and a universal 3D platform does not currently exist”
The paper relies on this without support.
Most 2D methods do not provide control of cell shape.
“However, most of these 2D methods do not provide control of cell shape, which determines biophysical cues affecting cell bioactivities in vivo.”
The paper relies on this without support.
Hepatocytes do not survive well under traditional 2D culture methods.
“Indeed, alternative methods to traditional 2D cell culture were developed for hepatocytes because they do not survive well under traditional 2D culture methods.”
The paper relies on this without support.
A major problem in the field is the lack of a standard approach in three-dimensional cultures.
“One major problem in the field is the lack of a standard approach in 3D cultures”
The paper relies on this without support.
Two-dimensional cell culture practice is standard and roughly comparable.
“whereas 2D cell culture practice is standard and roughly comparable.”
The paper relies on this without support.
Three-dimensional studies are becoming increasingly necessary to replicate the behaviors of some in vivo conditions.
“Classically, 2D studies have been favored, but 3D studies are becoming more necessary to replicate the behaviors of some in vivo conditions.”
The paper relies on this without support.
Accurate measurements of the forces exerted on the extracellular matrix are necessary for understanding physiological processes such as cancer or immune cell migration.
“Obtaining accurate measurements of forces exerted on the ECM are necessary to understand physiological processes such as cancer or immune cell migration.”
Claims other papers make about this one
Each is checked against this paper’s own text: does it say what the citing paper says it does?
The source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Cells grow on a fl at plastic surface as a monolayer, and this can result in loss of crucial cellular signaling pathways and changes in cell responses to stimuli [14 -16].”
From 3D In Vitro Model (R)evolution: Unveiling Tumor–Stroma Interactions (2021) · cites this paper as [14]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Nevertheless, 2D models are too simple and are unable to mimic the complexity and dynamic interactions of the TME.”
From 3D In Vitro Model (R)evolution: Unveiling Tumor–Stroma Interactions (2021) · cites this paper as [14]
Experimental — accuracy not yet measuredThe source backs part of the claim, not all of it — or backs it for a different population, condition or comparison.
In the citing paper
“Generally, 2D cell culture models lack cues provided by the extracellular matrix (ECM). These models are easy to maintain and cost-effective but do not allow the study of glia-neuron communication and crosstalk [119].”
From Alzheimer's Disease: Treatment Strategies and Their Limitations. (2022) · cites this paper as [119]
Experimental — accuracy not yet measuredThe cited source says what the claim attributes to it.
In the citing paper
“3D models can reproduce mechanical and biochemical cues that are crucial for cancer development, such as morphology, cell -cell/cell -ECM interactions, tissue stiffness, and speci fi c gradients [4,14,18].”
From 3D In Vitro Model (R)evolution: Unveiling Tumor–Stroma Interactions (2021) · cites this paper as [14]
Experimental — accuracy not yet measuredThe cited source says what the claim attributes to it.
In the citing paper
“The development of hepatocyte sandwich cultures (SCs), co-cultures with non-parenchymal cells and even more advanced 3-dimensional culturing techniques has revealed the importance of cell-cell and cell-matrix interactions for improving hepatic functions in liver models (Bachmann et al., 2015[2]; Duval et al., 2017[9]; Wei et al., 2018[45]).”
From Optimization of extracellular matrix for primary human hepatocyte cultures using mixed collagen-matrigel matrices (2023) · cites this paper as Duval et al., 2017[9]
Self-check
Percentages, sums and reported differences re-computed from the paper’s own numbers.
Each contradiction between two passages listed here was confirmed by an independent check.
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ 2D vs. 3D Cell Culture > Microfluidics.
“as a diagnostic tool in developing countries (95)”
§ 2D vs. 3D Cell Culture > References
“95. Salerno A, Guarnieri D, Iannone M, Zeppetelli S, Netti PA. Effect of micro-and macroporosity of bone tissue three-dimensional-poly(epsilon-cap- rolactone) scaffold on human mesenchymal stem cells invasion, proliferation, and differentiation in vitro.”
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ 2D vs. 3D Cell Culture > Abstract
“some important characteristics of cancer cells cannot be appropriately modeled in 2D cultures (22).”
§ 2D vs. 3D Cell Culture > References
“22. Choi SW, Yeh YC, Zhang Y, Sung HW, Xia Y. Uniform beads with controllable pore sizes for biomedical applications.”
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ 2D vs. 3D Cell Culture > Microfluidics.
“PDMS does not corrode or become affected by aqueous solutions used in cell culture, pressure, or temperature, and cells are able to survive or grow as a result (107).”
§ 2D vs. 3D Cell Culture > References
“107. Sun BK, Siprashvili Z, Khavari PA. Advances in skin grafting and treatment of cutaneous wounds.”
Rated minor: it would not touch a result — a value no result rests on, a p-value that stays on the same side of the significance level, or a conflict that is clerical, resolved elsewhere in the paper, or outside its own findings.
In the paper
§ 2D vs. 3D Cell Culture > Prefabricated scaffolds.
“gas foaming (96)”
§ 2D vs. 3D Cell Culture > References
“96. Sarvi F, Jain K, Arbatan T, Verma PJ, Hourigan K, Thompson MC, Shen W, Chan PPY. Cardiogen- esis of embryonic stem cells with liquid marble micro-bioreactor.”
Citations
Cites
Matrix elasticity directs stem cell lineage specification
Engler, A. J., Sen, S., Sweeney, H. L., & Discher, D. E. (2006) · Cell · doi:10.1016/j.cell.2006.06.044 · Cited by 13,915
The origins and the future of microfluidics
Whitesides, G. M. (2006) · Nature · doi:10.1038/nature05058 · Cited by 9,687
Tissue Cells Feel and Respond to the Stiffness of Their Substrate
Discher, D. E., Janmey, P., & Wang, Y.-L. (2005) · Science · doi:10.1126/science.1116995 · Cited by 6,382
Role of YAP/TAZ in mechanotransduction
Dupont, S., Morsut, L., Aragona, M., Enzo, E., Giulitti, S., Cordenonsi, M., Zanconato, F., Le Digabel, J., Forcato, M., Bicciato, S., et al. (2011) · Nature · doi:10.1038/nature10137 · Cited by 5,979
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Reconstituting organ-level lung functions on a chip
Huh, D., Matthews, B. D., Mammoto, A., Montoya-Zavala, M., Hsin, H. Y., & Ingber, D. E. (2010) · Science · doi:10.1126/science.1188302 · Cited by 4,250
Cell Shape, Cytoskeletal Tension, and RhoA Regulate Stem Cell Lineage Commitment
McBeath, R., Pirone, D. M., Nelson, C. M., Bhadriraju, K., & Chen, C. S. (2004) · Developmental Cell · doi:10.1016/s1534-5807(04)00075-9 · Cited by 4,182
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Taking Cell-Matrix Adhesions to the Third Dimension
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Cited by
Sorted by citations
Mechanisms of 3D cell migration
Yamada, K. M., & Sixt, M. (2019) · Nature Reviews Molecular Cell Biology · doi:10.1038/s41580-019-0172-9 · Cited by 906
Wound healing, fibroblast heterogeneity, and fibrosis
desJardins-Park, H. E., Mascharak, S., Griffin, M., Wan, D. C., & Longaker, M. T. (2022) · Cell stem cell · doi:10.1016/j.stem.2022.07.006 · Cited by 835
Alzheimer's Disease: Treatment Strategies and Their Limitations.
Passeri, E., Elkhoury, K., Morsink, M., Broersen, K., Linder, M., Tamayol, A., Malaplate, C., Yen, F. T., & Arab-Tehrany, E. (2022) · International Journal of Molecular Sciences · doi:10.3390/ijms232213954 · Cited by 689
“These models are easy to maintain and cost-effective but do not allow the study of glia-neuron communication and crosstalk [119].”
Background3D In Vitro Model (R)evolution: Unveiling Tumor–Stroma Interactions
Rodrigues, J., Heinrich, M. A., Teixeira, L. M., & Prakash, J. (2021) · Trends in Cancer · doi:10.1016/j.trecan.2020.10.009 · Cited by 405
“Cell interactions Deprived of cell–cell and cell–ECM interactions, no cell niches are created Cell junctions are common and allow cell communication [14]”
BackgroundConductive biomaterials for muscle tissue engineering
Dong, R., Ma, P. X., & Guo, B. (2019) · Biomaterials · doi:10.1016/j.biomaterials.2019.119584 · Cited by 387
Metadata
- Authors
- Duval, K., Grover, H., Han, L.-H., Mou, Y., Pegoraro, A. F., Fredberg, J., & Chen, Z.
- Affiliations
- Dartmouth College (US)
- Drexel University (US)
- University of Illinois Chicago (US)
- Harvard University (US)
- Corresponding author
- Chen, Zi
- Venue
- Physiology
- Publisher
- American Physiological Society
- Published
- 2017-07
- Volume
- 32
- Issue
- 4
- Pages
- 266-277
- ISSN
- 0886-1714 (ISSN-L)
- 1548-9213
- 1548-9221
- PMC
- PMC5545611
- OpenAlex
- W2626725994
- Cited by
- 1,849
- References
- 129
- Citations by year
- 2026: 184 · 2025: 240 · 2024: 251 · 2023: 252 · 2022: 241 · 2021: 268 · 2020: 234 · 2019: 127 · 2018: 46 · 2017: 4
- FWCI
- 9.03
- Type
- journal article
- Language
- en
- Open access
- hybrid
- doi.org
- Licence
- other-oa
- Locations
- Physiology (publishedVersion, other-oa)
- PubMed Central (publishedVersion)
- PubMed (publishedVersion)
- Funders
- Society in Science-Branco Weiss Fellowship
- HHS | NIH | National Cancer Institute (NCI) (U01CA202123)
- Topics
- Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Cell Biology › Cellular Mechanics and Interactions
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- 2D cell culture, 3D cell culture, tissue engineering, regenerative medicine, mechanical microenvironment, metabolic waste, physiological modeling, biomechanics
- MeSH terms
- Animals, Cell Differentiation, Cell Movement, Extracellular Matrix, Humans, Cell Culture Techniques, Tissue Engineering, Biomedical Research, Cell Proliferation
About these checks
Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.
A language model drew the claims from the paper on 2026-10-07, under version 3 of the claim definition (model deepseek-v4.1-flash, prompt extract-v3).
A second model labelled each claim on 2026-10-07 (model jev-1.13-20260917, definition revision gold-guideline-v0.2).
Cited sources are checked with a language model’s help (gpt-6-sol).
Statistics and arithmetic are recomputed, not judged.
Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.
What counts as a claim ›How we measure confidence ›How we check this ›
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