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Cited by 123D Printing in Biomedical ResearchLiver physiology and pathologyTissue Engineering and Regenerative Medicine

Fast formation and maturation enhancement of human liver organoids using a liver-organoid-on-a-chip

Open access
Byeon, Jae Hee, Jung, Da Jung, Han, Hyo-Jeong, Son, Woo-Chan, Jeong, Gi Seok·2024·Frontiers in Cell and Developmental Biology
DOI 10.3389/fcell.2024.1452485PMC PMC11349704
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Abstract

Background: Spatial and functional hepatic zonation, established by the heterogeneous tissue along the portal–central axis of the liver, is important for ensuring optimal liver function. Researchers have attempted to develop reliable hepatic models to mimic the liver microenvironment and analyze liver function using hepatocytes cultured in the developed systems. However, mimicking the liver microenvironment in vitro remains a great challenge owing to the lack of perfusable vascular networks in the model systems and the limitation in maintaining hepatocyte function over time. Methods: In this study, we established a microphysiological system that operated under continuous flush medium flow, thereby allowing the supply of nutrients and oxygen to liver organoids and the removal of waste and release of cytokines therefrom, similar to the function of blood vessels. Results: The application of microphysiological system to organoid culture was advantageous for reducing the differentiation time and enhancing the functional maturity of human liver organoid. Conclusion: Hence, our microphysiological culture system might open the possibility of the miniaturized liver model system into a single device to enable more rational in vitro assays of liver response.

AI snapshot

Written from the paper’s full text.

TL;DR

The paper describes a liver-organoid-on-a-chip microphysiological system (MPS) with continuous medium flow to culture human liver organoids (hLOs). Compared with conventional Matrigel droplet static culture, the MPS shortened the expansion/differentiation period to approximately 11 days and enhanced hepatocyte-like maturation and function.

Key findings

  • hLOs in the MPS completed expansion and differentiation in approximately 11 days total, whereas Matrigel droplet cultures typically needed a minimum of 21 days or longer to fully differentiate.
  • During expansion, CD133 expression was higher in MPS hLOs than in droplet cultures; EpCAM, SOX9, and KRT19 were expressed under both static and flow conditions, with no significant differences in immunofluorescence for SOX9, LGR5, KRT19, and EpCAM.
  • After differentiation, albumin and CYP3A4 mRNA were highly expressed under flow versus static; A1AT expression in MPS increased by approximately 10% over static; CYP450s were higher in fluidic systems; bile acid-related metabolites were secreted in high amounts on day 4.
  • MPS organoids showed less size/morphological heterogeneity and clearer polygonal structure/hepatocyte marker expression (ZO-1, HNF4α, MRP2, albumin), with albumin highly accumulated inside fluidic-system organoids; MRP2 was scarcer under static.

Methods

  • PDMS microfluidic device replicated from an SU-8 mold, bonded to cover glass, with 29 microwells; organoids seeded with 50% growth factor-reduced Matrigel via centrifugal force and cultured under continuous medium flow controlled by a yarn flow resistor.
  • Human adjacent non-tumor liver tissue from a consented patient at Asan Medical Center; liver ductal cells isolated, hepatic bipotent stem cells expanded, and hLOs formed. Static Matrigel droplet cultures served as comparator.
  • Analyses: qRT-PCR (normalized to GAPDH, 2^-ΔΔCT), immunofluorescence/whole-mount tissue clearing, flow cytometry for A1AT, ELISA for albumin, LC-MS/MS for bile acids, unpaired Student’s t-test with P<0.05; data mean ± S.D. of triplicates.

Limitations

  • The paper does not present a dedicated limitations section. It notes general MPS hurdles in the field: complex procedures requiring specialized equipment/materials and absence of organoid size control.
  • Evident from the text: experiments used adjacent non-tumor liver tissue from a single consented patient, so donor variability is not addressed.
  • Evident from the text: some comparisons are qualitative (e.g., immunofluorescence morphology/marker clarity), and the MPS culture medium required concentration before ELISA due to larger culture volume.

Model-written summary. Check it against the paper before relying on it.

Integrity signals

Registry

Crossref · 10.3389/fcell.2024.1452485

Cites

37 of 38 references verified1 citation errors · 0 not checkable

Statistics

Nothing to recompute.

Claims the paper makes

21 of 34 cited sources checked · 12 supported · 0 flagged · accuracy not yet measured

  • Premisecitation

    There remain hurdles to overcome in MPSs, including an absence of organoid size control.

    “These challenges include complex procedure requiring specialized equipment and materials, as well as an absence of organoid size control (Balakrishnan et al., 2015;Schuster et al., 2020).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Balakrishnan et al., 2015 source

      The Introduction discusses specialized systems for live-cell imaging and perfusion, and the Results and Discussion describes cells grown on microfabricated scaffolds. Neither passage addresses organoids or control of their size; searches for “organoid size control” and the cited sentence’s description of the challenge found no such discussion in our copy. (LLM, conf=0.76)

      Experimental — accuracy not yet measured
    • cites Schuster et al., 2020 source

      The Introduction says some body-on-a-chip systems require complex or expensive robotic equipment, supporting the equipment-related challenge. It also says existing microfluidic chambers can be too shallow for large organoids, but neither the Introduction nor the Results passage on chamber design identifies an absence of organoid size control. The paper discusses accommodating and measuring organoid size, not controlling it. (LLM, conf=0.84)

      Experimental — accuracy not yet measured
  • Premisecitation

    Hepatic zonation along the portal-central axis is important for optimal liver function.

    “Spatial and functional hepatic zonation, established by the heterogeneous tissue along the portal-central axis of the liver, is important for ensuring optimal liver function.”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Tsukada et al., 2013 source
    • cites Ben-Moshe and Itzkovitz, 2019 source
  • Premisecitation

    Blood flow from the periportal to the perivenous zones along the lobular axis creates spatial gradients of oxygen, metabolites, and nutrients, which benefit hepatocytes and other cells.

    “Blood flow from the periportal to the perivenous zones along the lobular axis creates spatial gradients of oxygen, metabolites, and nutrients, which benefit hepatocytes and other cells (Tsukada et al., 2013;Ben-Moshe and Itzkovitz, 2019).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Tsukada et al., 2013 source
    • cites Ben-Moshe and Itzkovitz, 2019 source

      The Introduction (page not specified) says blood flows from portal nodes toward central veins, generating gradients of oxygen and nutrients, and links the graded environment to hepatocyte zonation. It names hormones rather than metabolites as a blood-flow-generated gradient, and does not say these gradients benefit other cell types. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
  • Premisecitation

    Depending on the hepatic cell position in the hepatic lobule and the corresponding differences in exposure to oxygen and nutrient gradients, heterogeneity and functional plasticity arose among the hepatic cells.

    “Depending on the hepatic cell position in the hepatic lobule and the corresponding differences in exposure to oxygen and nutrient gradients, heterogeneity and functional plasticity arose among the hepatic cells (Tonon et al., 2019).”
    Quote in the paperSubstantial claim1 of 1 sources checked
    • cites Tonon et al., 2019 source
  • Findingreasoning

    Self-assembled hLOs cultured in the MPS displayed phenotypic and functional enhancements over those formed using the conventional droplet culture method.

    “The self-assembled hLOs cultured in the MPS displayed phenotypic and functional enhancements over those formed using the conventional droplet culture method, indicating that the MPS had additional beneficial effects on the maturity of the organoids as it provided a more invivo-like cellular microenvironment.”
    Quote in the paperSubstantial claim
  • Classified as probably not a substantial claim. 23
  • May combine two claims. 8

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

    Premisecitation

    Physiological relevant in vitro models provide a promising platform for modelling the physiochemical conditions of the hepatic microenvironment.

    “Physiological relevant in vitro models provide a promising platform for modelling the physiochemical conditions of the hepatic microenvironment (Janani and Mandal, 2021;Kostrzewski et al., 2021;Paradiso et al., 2023).”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites Janani and Mandal, 2021 source
    • cites Kostrzewski et al., 2021 source
    • cites Paradiso et al., 2023 source

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

    Premisecitation

    The organoid could recapitulate several liver diseases, including fatty liver disease, cirrhosis, and monogenic metabolic diseases.

    “Furthermore, the organoid could recapitulate several liver diseases, including fatty liver disease, cirrhosis, and monogenic metabolic diseases (Guan et al., 2017;Wang et al., 2020;Guan et al., 2021).”
    Quote in the paperNot a substantial claim1 of 3 sources checked
    • cites Guan et al., 2017 source
    • cites Wang et al., 2020 source
    • cites Guan et al., 2021 source

      The Introduction and Results report that the organoids reproduce ARPKD liver pathology, a genetic disease involving fibrosis. The Results section on similarities with other forms of fibrosis finds gene-expression similarities with cirrhotic liver and non-alcoholic steatohepatitis, but does not show that the organoids recapitulate cirrhosis or fatty liver disease as distinct diseases. The paper also does not establish a model of monogenic metabolic diseases generally. (LLM, conf=0.89)

      Experimental — accuracy not yet measured

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

    Premisecitation

    The liver organoid has self-renewal capability, a key proliferative property for its long-term culture and clonal expansion from single cells with sustained stemness.

    “The liver organoid has selfrenewal capability, a key proliferative property for its long-term culture and clonal expansion from single cells with sustained stemness (Huch et al., 2013;Hu et al., 2018).”
    Quote in the paperNot a substantial claim1 of 2 sources checked
    • cites Huch et al., 2013 source
    • cites Hu et al., 2018

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

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

    Premisecitation

    Integration of the perfusion channel for dynamic culture has allowed the recapitulation of in vivo conditions, leading to spatiotemporal distribution of oxygen, nutrients and metabolic wastes.

    “The Integration of the perfusion channel for dynamic culture has allowed the recapitulation of in vivo conditions, leading to spatiotemporal distribution of oxygen, nutrients and metabolic wastes (Yu et al., 2017;Lekkala et al., 2024).”
    Quote in the paperSubstantial claim1 of 2 sources checked
    • cites Yu et al., 2017

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Lekkala et al., 2024 source

      The Discussion (page not provided) says perfusion enhanced oxygen and nutrient diffusion and waste removal, while the Introduction describes static spheroids as having limited in vivo-like tissue structure. The paper does not establish that the perfusion plate recapitulates in vivo conditions or demonstrate a spatiotemporal distribution of those substances. (LLM, conf=0.84)

      Experimental — accuracy not yet measured

  • Classified as probably not a substantial claim.

    Premisecitation

    There remain hurdles to overcome in MPSs, including complex procedure requiring specialized equipment and materials.

    “These challenges include complex procedure requiring specialized equipment and materials, as well as an absence of organoid size control (Balakrishnan et al., 2015;Schuster et al., 2020).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Balakrishnan et al., 2015 source

      The Introduction says these cell-culture studies require specialized systems and describes conventional perfusion-pump setups as cumbersome. It does not establish that MPS procedures generally require specialized materials, so the broader claim exceeds what this paper supports. The separate point about organoid size control is not assessed here. (LLM, conf=0.82)

      Experimental — accuracy not yet measured
    • cites Schuster et al., 2020 source

  • Classified as probably not a substantial claim.

    Premisecitation

    Three-dimensional cell cultures, especially organoids, have been proposed for mimicking the architecture and genetic expression of native human liver tissue.

    “To overcome the limitations of these 2D in vitro models, three-dimensional (3D) cell cultures-especially organoids-have been proposed for mimicking the architecture and genetic expression of native human liver tissue (Akbari et al., 2019;Sorrentino et al., 2020).”
    Quote in the paperSubstantial claim2 of 2 sources checked
    • cites Akbari et al., 2019 source
    • cites Sorrentino et al., 2020 source

      The Results section, “PEG hydrogels allow derivation and culture of human liver organoids” (page unavailable), reports 3D human liver organoids that express a progenitor marker and can differentiate into hepatocyte-like cells. This supports the use of organoids to model aspects of human liver cells, but the cited passages do not establish that they mimic the architecture or overall gene-expression profile of native human liver tissue. (LLM, conf=0.83)

      Experimental — accuracy not yet measured

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

    Premisecitation

    The liver is a spatially and functionally heterogeneous organ that contains branches of the hepatic portal vein, hepatic artery, and biliary tree across its lobules.

    “The liver is a spatially and functionally heterogeneous organ that contains branches of the hepatic portal vein, hepatic artery, and biliary tree across its lobules (Ben-Moshe et al., 2019;Inverso et al., 2021).”
    Quote in the paperNot a substantial claim2 of 2 sources checked
    • cites Ben-Moshe et al., 2019 source

      The Introduction (page unavailable) describes lobules and spatial heterogeneity in hepatocyte gene expression, while the Discussion describes functional differences between zones. The paper mentions blood flowing from portal nodes but does not describe branches of the hepatic portal vein, hepatic artery, and biliary tree across lobules. (LLM, conf=0.86)

      Experimental — accuracy not yet measured
    • cites Inverso et al., 2021 source

      The Introduction (page unavailable) describes blood entering the liver through the hepatic artery and portal vein and describes heterogeneity along the lobule’s portal-to-central axis. The Discussion (page unavailable) also describes spatial and molecular heterogeneity of the liver endothelium. The passages do not describe branches of a biliary tree across the lobules, so that part of the attributed claim is unsupported. (LLM, conf=0.85)

      Experimental — accuracy not yet measured

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

    Premisecitation

    Self-renewing liver organoids demonstrating genetic stability during long-term culture were reported in 2015.

    “Self-renewing liver organoids demonstrating genetic stability during long-term culture were reported in 2015 (Huch et al., 2015).”
    Quote in the paperNot a substantial claim1 of 1 sources checked
    • cites Huch et al., 2015 source

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

    Premisecitation

    Recently, stem cell-based 3D culture methods have been developed for the formation of organoids.

    “Recently, stem cell-based 3D culture methods have been developed for the formation of organoids (Sato et al., 2011;Fatehullah et al., 2016;Boonekamp et al., 2019).”
    Quote in the paperNot a substantial claim2 of 3 sources checked
    • cites Sato et al., 2011 source
    • cites Fatehullah et al., 2016 source
    • cites Boonekamp et al., 2019 source

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

    Findingreasoning

    Through the process of diffusion, the soluble factors, growth factors, nutrients, and oxygen were indirectly supplied to the hLOs in the microwells.

    “Hence, through the process of diffusion, the soluble factors, growth factors, nutrients, and oxygen were indirectly supplied to the hLOs in the microwells.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    The application of microphysiological system to organoid culture was advantageous for reducing the differentiation time and enhancing the functional maturity of human liver organoid.

    “The application of microphysiological system to organoid culture was advantageous for reducing the differentiation time and enhancing the functional maturity of human liver organoid.”
    Quote in the paperSubstantial claim

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

    Findingnone given

    The microfluidic platform provides an easy-to-use, low-cost, and single-step 3D microfluidic device for culturing human liver organoids.

    “Our microfluidic platform provides an easy-to-use, lowcost, and single-step 3D microfluidic device for culturing human liver organoids.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Findingown data

    Oxygen and nutrient diffusion is less limited in MPS due to its relatively thin layer of Matrigel, resulting in a narrow range of variation in the size and morphological heterogeneity of the organoids.

    “In contrast, oxygen and nutrient diffusion is less limited in MPS due to its relatively thin layer of Matrigel, resulting in a narrow range of variation in the size and morphological heterogeneity of the organoids (Figure 2B; Supplementary Figure S3).”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Both albumin and CYP3A4 markers were highly expressed under the flow condition compared with the static condition.

    “Both markers were highly expressed under the flow condition compared with under the static condition.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Findingown data

    Morphological heterogeneity was observed in some hLOs in the droplet, and these changes occurred irregularly during differentiation.

    “Morphological heterogeneity was observed in some hLOs in the droplet, and these changes occurred irregularly during differentiation.”
    Quote in the paperNot a substantial claim

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

    Findingown data

    During the expansion process, the hLOs in the Matrigel droplet were observed to be varied in size, with those near the boundary of the droplet being larger than those at the core region.

    “During the expansion process, the hLOs in the Matrigel droplet were observed to be varied in size, with those near the boundary of the droplet being larger than those at the core region (Supplementary Figure S2).”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Interpretationreasoning

    Liver organoids are a promising cell source for in vitro studies on the developmental biology and pathophysiology of the liver and hepatic system.

    “Results have shown liver organoids to be a promising cell source for in vitro studies on the developmental biology and pathophysiology of the liver and hepatic system.”
    Quote in the paperSubstantial claim

  • May combine two claims.

    Interpretationreasoning

    The MPS had additional beneficial effects on the maturity of the organoids as it provided a more in vivo-like cellular microenvironment.

    “The self-assembled hLOs cultured in the MPS displayed phenotypic and functional enhancements over those formed using the conventional droplet culture method, indicating that the MPS had additional beneficial effects on the maturity of the organoids as it provided a more invivo-like cellular microenvironment.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    The system is designed to localize organoids at approximately the same position along the fabricated microfluidic channel, so organoids can be exposed to similar environmental conditions.

    “Moreover, this system is designed to localize organoids at approximately the same position along the fabricated microfluidic channel, so organoids can be exposed to similar environmental conditions.”
    Quote in the paperSubstantial claim

  • Classified as probably not a substantial claim.

    Premisecitation

    The liver is organized into units known as lobules.

    “In terms of its anatomical structure, the liver is organized into units known as lobules (Trefts et al., 2017).”
    Quote in the paperSubstantial claim0 of 1 sources checked
    • cites Trefts et al., 2017 source

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

    Premisecitation

    The spatial heterogeneity in the liver serves a wide variety of physiological functions, including xenobiotic metabolism, protein and bile synthesis, and the dynamics of hepatic regeneration.

    “The spatial heterogeneity in the liver serves a wide variety of physiological functions, including xenobiotic metabolism, protein and bile synthesis, and the dynamics of hepatic regeneration (Li and Chiang, 2014;Tanami et al., 2017).”
    Quote in the paperNot a substantial claim0 of 2 sources checked
    • cites Li and Chiang, 2014

      Couldn’t be matched to a paper in Crossref, so it can’t be checked.

    • cites Tanami et al., 2017 source

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

    Study design and conductown data

    Experiments were performed in triplicate.

    “Data were expressed as mean ± S.D. of triplicate experiments.”
    Quote in the paperNot a substantial claim

  • May combine two claims.

    Interpretationown data

    Overall, hepatocytes produced under the dynamic medium flow condition functionally outperformed those from the conventional droplet culture in albumin production, upregulation of CYP450s expression, and accumulation of bile acids.

    “Overall, hepatocytes produced under the dynamic medium flow condition functionally outperformed those from the conventional droplet culture in albumin production, upregulation of CYP 450s expression, and accumulation of bile acids.”
    Quote in the paperSubstantial claim

  • Probably not a substantial claim: may not pass S3 (load-bearing), S4 (based).

    Interpretationreasoning

    CYP3A4 was activated by endogenous bile acid for detoxification of the latter.

    “We could presume that CYP3A4 was activated by endogenous bile acid for detoxification of the latter.”
    Quote in the paperNot a substantial claim

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

    Study design and conductnone given

    A1AT expression was determined based on samples stained with the isotype controls.

    “A1AT expression was determined based on samples stained with the isotype controls.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    According to previous reports, isolated healthy ductal cells (or LGR5-positive liver cells post-damage induction) could self-organize into 3D structures that sustained long-term expansion as adult ductal progenitor cells while retaining their ability to differentiate into functional hepatocyte-like cells in vitro.

    “According to previous reports, isolated healthy ductal cells (or LGR5-positive liver cells post-damage induction) could self-organize into 3D structures that sustained long-term expansion as adult ductal progenitor cells while retaining their ability to differentiate into functional hepatocyte-like cells in vitro.”
    Quote in the paperNot a substantial claim

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

    Premisecitation

    Self-organizing 3D structures are formed by embedding tissue-resident stem cells in Matrigel and then culturing them in media supplemented with growth factors based on the endogenous stem cell niche.

    “These selforganizing 3D structures are formed by embedding tissueresident stem cells in Matrigel and then culturing them in media supplemented with growth factors based on the endogenous stem cell niche (Broutier et al., 2016).”
    Quote in the paperNot a substantial claim0 of 1 sources checked
    • cites Broutier et al., 2016 source

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

    Study design and conductown data

    The calibration range for each lipid was 0.1-10,000 nM (r2 ≥ 0.99).

    “The calibration range for each lipid was 0.1-10,000 nM (r 2 ≥ 0.99).”
    Quote in the paperNot a substantial claim

  • Classified as probably not a substantial claim.

    Study design and conductnone given

    Statistical significance was considered at a P-value of less than 0.05 between groups.

    “A P-value of less than 0.05 between groups was considered statistically significant.”
    Quote in the paperSubstantial claim
29 of 29

  • The paper relies on this without support.

    Mimicking the liver microenvironment in vitro remains a great challenge owing to the lack of perfusable vascular networks in the model systems and the limitation in maintaining hepatocyte function over time.

    “However, mimicking the liver microenvironment in vitro remains a great challenge owing to the lack of perfusable vascular networks in the model systems and the limitation in maintaining hepatocyte function over time.”

  • The paper relies on this without support.

    Most conventional in vitro liver models have been unable to fully replicate the complex tissue architecture, regenerative capacity, and functions of the liver.

    “Most of the conventional in vitro liver models have been unable to fully replicate the complex tissue architecture, regenerative capacity, and functions of this organ.”

  • The paper relies on this without support.

    It has been challenging to maintain long-term cultures and develop the phenotype and functions of fully mature hepatocytes.

    “However, it has been challenging to maintain long-term cultures and develop the phenotype and functions of fully mature hepatocytes.”

  • The paper relies on this without support.

    Conventional droplet-based culture conditions do not possess the unique hepatic microenvironment required to preserve biochemical gradients, such as those of nutrients, oxygen tension, and metabolites.

    “In particular, the conventional droplet-based culture conditions do not possess the unique hepatic microenvironment required to preserve biochemical gradients, such as those of nutrients, oxygen tension, and metabolites.”

  • The paper relies on this without support.

    Many current in vitro liver models are limited by their lack of proper hepatic functions, the loss of the hepatic microenvironment, and the short lifespan of the hepatic cells in culture.

    “However, many current in vitro liver models are limited by their lack of proper hepatic functions, the loss of the hepatic microenvironment, and the short lifespan of the hepatic cells in culture.”

  • The paper relies on this without support.

    Matrigel-based droplet culture cannot provide nutrients and oxygen exchange and limits the amount of metabolic waste being removed from the interior of the organoids.

    “However, the Matrigel-based droplet culture cannot provide nutrients and oxygen exchange and limits the amount of metabolic waste being removed from the interior of the organoids.”

  • The paper relies on this without support.

    Owing to limited diffusion of oxygen and nutrients into their core, organoids of irregular size were generated, and differences in the expansion and differentiation stages occurred.

    “Owing to the limited diffusion of oxygen and nutrients into their core, organoids of irregular size were generated, and differences in the expansion and differentiation stages occurred.”
7 of 7

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.

  • MPS culture stages have conflicting durations

    Minor

    The results say MPS organoids spent 5 days in expansion medium and 6 days in differentiation medium. Figure 1 instead specifies 4 days in expansion medium, 2 days of BMP7 treatment, and 5 days in differentiation medium; the methods also give a 5-day differentiation period.

1 of 1

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Related

Authors
Byeon, J. H., Jung, D. J., Han, H.-J., Son, W.-C., & Jeong, G. S.
Affiliations
Asan Medical Center (KR)
Corresponding author
  • Son, Woo-Chan
  • Jeong, Gi Seok
ORCID
Venue
Frontiers in Cell and Developmental Biology
Publisher
Frontiers Media SA
Published
2024-08-14
Volume
12
Pages
1452485-1452485
ISSN
2296-634X (ISSN-L)
DOI
10.3389/fcell.2024.1452485
PMC
PMC11349704
OpenAlex
W4401590434
Cited by
12
References
36
Citations by year
2026: 5 · 2025: 7
FWCI
1.31
Type
journal article
Language
en
Open access
Licence
Locations
  • Frontiers in Cell and Developmental Biology (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (submittedVersion, cc-by)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion, cc-by-sa)
Topics
  • Physical Sciences › Engineering › Biomedical Engineering › 3D Printing in Biomedical Research
  • Health Sciences › Medicine › Hepatology › Liver physiology and pathology
  • Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
Keywords
liver organoids, organ-on-a-chip, microphysiological systems, continuous perfusion, hepatic zonation, hepatocyte differentiation, organoid maturation, liver microenvironment

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

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