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Cited by 1,8493D Printing in Biomedical ResearchCellular Mechanics and InteractionsTissue Engineering and Regenerative Medicine

Modeling physiological events in 2D vs. 3D cell culture

Open access
Duval, Kayla, Grover, Hannah, Han, Li-Hsin, Mou, Yongchao, Pegoraro, Adrian F., Fredberg, Jeffery, Chen, Zi·2017·Physiology
DOI 10.1152/physiol.00036.2016PMC PMC5545611
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Abstract

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

Integrity signals

Registry

Crossref · 10.1152/physiol.00036.2016

Cites

129 of 131 references verified0 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

44 of 86 cited sources checked · 18 supported · 1 flagged · accuracy not yet measured

  • Premisecitation

    Cell differentiation, migration, growth, and mechanics are influenced by the biochemical and biomechanical microenvironment.

    “These behaviors include cell differentiation, migration, growth, and mechanics, all of which are impacted by their biochemical and biomechanical microenvironment (57).”
    Quote in the paperSubstantial claim1 of 1 sources checked
  • Premisecitation

    Some important characteristics of cancer cells cannot be appropriately modeled in 2D cultures.

    “For instance, some important characteristics of cancer cells cannot be appropriately modeled in 2D cultures (22).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [22] source

      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)

      Experimental — accuracy not yet measured
  • Premisecitation

    Increasing the dimensionality of extracellular matrix around cells from 2D to 3D can significantly impact cell proliferation, differentiation, mechano-responses, and survival.

    “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).”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [2] source
    • cites [10] source

      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)

      Experimental — accuracy not yet measured
    • cites [41] source

      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)

      Experimental — accuracy not yet measured
  • Premisecitation

    The sandwich culture method has long been demonstrated to produce hepatocyte cell cultures where morphology and function more accurately reproduce in vivo behavior.

    “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).”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [9] source
    • cites [29] source

      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)

      Experimental — accuracy not yet measured
    • cites [34] source
  • Premisecitation

    Cells in the body perform bioactivities in response to stimulation from a highly complex, 3D microenvironment.

    “Cells in our body perform bioactivities in response to the stimulation from a highly complex, 3D microenvironment (15,98,112).”
    Quote in the paperSubstantial claim2 of 3 sources checked
    • cites [15] source
    • cites [98] source

      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)

      Experimental — accuracy not yet measured
    • cites [112] source
  • May combine two claims. 16
  • Classified as probably not a substantial claim. 14

  • May combine two claims.

    Premisecitation

    PDMS wells with triangular pyramid shape opening promoted growth of spheroids, with some control over size achievable.

    “PDMS wells with a triangular pyramid shape opening promoted the growth of spheroids, with some control over the size achievable.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [110] source

      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.

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Novel 3D cell culture platforms are being created to better mimic in vivo conditions and are sometimes called spheroid or organoid culture.

    “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).”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [37] source
    • cites [61] source

      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)

      Experimental — accuracy not yet measured
    • cites [84] source

  • May combine two claims.

    Premisecitation

    Wan et al. demonstrated that cells preferred to round up on smooth-surfaced PLLA substrates and stretch on rough surfaces.

    “They demonstrated that cells preferred to round up on smooth-surfaced PLLA substrates and stretch on rough surfaces.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [113] source

      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)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Torisawa et al. created a chip system utilizing microfluidics that allows spheroids to grow in a controlled environment for long-term culture.

    “Torisawa et al. (110) created a novel chip system utilizing microfluidics that allows spheroids to grow in a controlled environment for long-term culture.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [110] source

      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)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Dunn et al. found that sandwich culture maintained proper albumin production rate for at least 6 weeks, whereas the one-gel system had rates tenfold lower than normal by the end of the first week.

    “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).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [29] source

      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)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Cell growth in 2D monolayers allows access to a similar amount of nutrients and growth factors present in the medium, resulting in homogenous growth and proliferation.

    “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).”
    Quote in the paperNot a substantial claim1 of 1 sources checked

  • May combine two claims.

    Premisecitation

    In 2D cultures, there are decreases in transcriptional genes and rapid changes in morphology.

    “In 2D cultures, there are decreases in transcriptional genes and rapid changes in morphology (29,34).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites [29] source

      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)

      Experimental — accuracy not yet measured
    • cites [34] source

  • May combine two claims.

    Premisecitation

    Wan et al. experiments revealed that pseudopods extending from cells could encroach into pits in culture surfaces; less rigid pseudopods could act as anchors and influence migration patterns, providing contact guidance.

    “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).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [113] source

      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)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Surfaces of aggregated spheroids have highest levels of proliferation, whereas interior of 3D cell bodies possesses highest number of quiescent or necrotic cells.

    “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).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [31] source

      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)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Hsiao et al. used these plates as a high throughput method for testing anticancer drug sensitivities on H9 human epithelial carcinoma cells.

    “They continued on to use these plates as a high throughput method for testing anticancer drug sensitivities on H9 human epithelial carcinoma cells.”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites [55] source

      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)

      Experimental — accuracy not yet measured

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Biopolymers from non-mammal tissues such as alginate and chitosan are also used for making 3D scaffolds.

    “Biopolymers from non-mammal tissues, such as alginate and chitosan, are also used for making 3D scaffolds (23,63,68,84).”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites [23] source

      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)

      Experimental — accuracy not yet measured
    • cites [63] source
    • cites [68] source

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Hsiao et al. designed a plate using micro-ring structures to stabilize drops, allowing 14-day culture of prostate cancer cells.

    “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.”
    Quote in the paperNot a substantial claim1 of 1 sources checked

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Among most commonly used natural polymers are hyaluronic acid, gelatin, collagen, and chondroitin sulfate.

    “Among the most commonly used natural polymers are hyaluronic acid, gelatin, collagen, and chondroitin sulfate (14, 108).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites [14] source

      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)

      Experimental — accuracy not yet measured
    • cites [108] source

  • May combine two claims.

    Premisecitation

    In mouse embryonic stem cells, OCT4 levels decreased in both two-dimensional and three-dimensional cultures, indicating a loss of pluripotency, whereas NES and BRACHY-T markers increased.

    “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.”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [91] source

      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)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    Results from 2D and 3D spheroid culture indicated spheroids were more resistant to 5-fluorouracil, whereas 2D cultures were more resistant to tirapazamine.

    “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).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites [111] source

      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)

      Experimental — accuracy not yet measured

  • May combine two claims.

    Premisecitation

    LeCluyse et al. found that hepatocyte polarity was affected on single-layer gels; sandwich culture produced normal distribution of apical membrane markers at the canalicular membrane and well-established bile canaliculi.

    “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).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [73] source

  • May combine two claims.

    Interpretationcitation

    Micro-patterning and type of surface both affected the rate of differentiation of cells.

    “The results indicated that the micro-patterning and type of surface both affected the rate of differentiation of cells (18).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites [18] source

  • Probably not a substantial claim: may not pass S2 (checkable).

    Interpretationreasoning

    3D platforms are likely to become an increasingly attractive alternative to 2D cell culture as technology develops to enable a wider range of processes.

    “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.”
    Quote in the paperNot a substantial claim
    • Rests on an assumption: 2D cell culture approaches can still recapitulate in vivo behavior for many bioactivities.

  • Probably not a substantial claim: may not pass S3 (load-bearing). May combine two claims.

    Premisecitation

    Micropatterning can be used to pattern areas or create microwells for controlled growth of spheroids.

    “Micropatterning can also be used to pattern areas or to create “microwells” for the controlled growth of spheroids (i.e., agarose) (35).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [35] source

  • Classified as probably not a substantial claim.

    Interpretationnone given

    These three-dimensional methods allow investigation of cell and tissue behavior in a more physiologically relevant environment.

    “These methods allow for a more physiologically relevant environment to investigate cell and tissue behavior.”
    Quote in the paperSubstantial claim
    • Rests on an assumption: Three-dimensional studies are becoming increasingly necessary to replicate the behaviors of some in vivo conditions.

  • Probably not a substantial claim: may not pass S2 (checkable), S4 (based).

    Interpretationnone given

    Different cell types are likely to be better matched to particular three-dimensional culture systems.

    “different cell types will most likely be better matched for certain 3D systems.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing).

    Premisecitation

    Microwells made out of non-adhesion promoting material such as agarose can be used to grow several arrays of spheroids at once.

    “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).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites [86] source

  • Probably not a substantial claim: may not pass S4 (based). May combine two claims.

    Interpretationnone given

    In some situations, two-dimensional studies are more feasible and can produce meaningful results.

    “In some cases, 2D studies are more feasible, and the results can be meaningful.”
    Quote in the paperNot a substantial claim

  • Probably not a substantial claim: may not pass S2 (checkable).

    Interpretationreasoning

    Whether a two-dimensional or a three-dimensional culture method is best suited depends on the situation and the questions being addressed.

    “In conclusion, the situation and questions at hand will determine whether a 2D or 3D culture method is best suited.”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim. May combine two claims.

    Interpretationreasoning

    In other situations, three-dimensional studies yield more accurate results and can be carried out together with two-dimensional studies.

    “However, in other cases, 3D studies will yield more accurate results and can be done in combination with 2D studies.”
    Quote in the paperSubstantial claim
25 of 25

  • 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.”
9 of 9

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?

  • Cells grown on flat plastic as a monolayer can lose crucial cellular signaling pathways and change responses to stimuli.

    “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]

    The Considerations section (page unavailable) describes conventional 2D culture on flat glass or polystyrene surfaces and says 2D-induced polarity may alter how some cells sense soluble factors and other cues. The paper also reports altered cellular behavior in 2D culture, but the passages do not establish a loss of crucial cellular signaling pathways. (LLM, conf=0.85)

    Experimental — accuracy not yet measured

  • 2D models are too simple and unable to mimic the complexity and dynamic interactions of the TME.

    “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]

    The Abstract says 2D cultures can fail to model some cancer-cell characteristics, and the Motion and Migration section describes interactions and signaling observed in 3D but not 2D. The claim overstates this as a categorical inability to mimic the tumor microenvironment: the Abstract also says 2D cultures can recapitulate many in vivo behaviors. Page numbers were not supplied. (LLM, conf=0.88)

    Experimental — accuracy not yet measured

  • 2D cell culture models lack cues provided by the extracellular matrix and do not allow the study of glia-neuron communication and crosstalk.

    “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]

    The Considerations and Extracellular Matrix sections describe how conventional 2D cultures lack some in vivo biophysical and ECM cues, and the Considerations section calls them simple and efficient. The paper does not say that 2D cultures prevent the study of glia–neuron communication or crosstalk; its Techniques section discusses hippocampal neuron culture without addressing that claim. No page numbers are available in our copy. (LLM, conf=0.87)

    Experimental — accuracy not yet measured

  • 3D models can reproduce mechanical and biochemical cues crucial for cancer development, such as morphology, cell-cell/cell-ECM interactions, tissue stiffness, and specific gradients.

    “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 measured

  • Cell-cell and cell-matrix interactions are important for improving hepatic functions in liver models.

    “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.

  • Diagnostic-tool claim cites a scaffold study

    Minor

    Reference 95 is identified as a study of bone-scaffold porosity and stem cells, rather than microfluidic diagnostic tools.

  • Cancer-cell claim cites a paper on porous beads

    Minor

    The citation attached to the claim about cancer cells in 2D culture is listed as a paper on beads with controllable pore sizes.

  • PDMS claim cites a skin-grafting review

    Minor

    The cited reference 107 is identified as a paper on skin grafting and wound treatment, not PDMS behavior in cell-culture devices.

  • Gas-foaming citation identifies a bioreactor study

    Minor

    The reference given for scaffold fabrication by gas foaming is listed as a study of embryonic stem-cell cardiogenesis using a liquid-marble micro-bioreactor.

4 of 4

Cites

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Related

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
ORCID
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
DOI
10.1152/physiol.00036.2016
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
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.

Claims read by deepseek-v4.1-flash under definition version 3 (extract-v3) on 2026-10-07.

Classified by jev-1.13-20260917 (gold-guideline-v0.2) on 2026-10-07.

Cited sources are checked by 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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