Extracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cells
Abstract Background Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells (HLCs) can provide a valuable in vitro model for disease modelling and drug development. However, generating HLCs with characteristics comparable to hepatocytes in vivo is challenging. Extracellular matrix (ECM) pla...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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BMC
2023-11-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | https://doi.org/10.1186/s13287-023-03542-x |
| _version_ | 1797637036756172800 |
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| author | Faiza Farhan Manjari Trivedi Priscilla Di Wu Wei Cui |
| author_facet | Faiza Farhan Manjari Trivedi Priscilla Di Wu Wei Cui |
| author_sort | Faiza Farhan |
| collection | DOAJ |
| description | Abstract Background Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells (HLCs) can provide a valuable in vitro model for disease modelling and drug development. However, generating HLCs with characteristics comparable to hepatocytes in vivo is challenging. Extracellular matrix (ECM) plays an important role in supporting liver development and hepatocyte functions, but their impact on hepatocyte differentiation and maturation during hPSC differentiation remains unclear. Here, we investigate the effects of two ECM components—Matrigel and type I collagen on hepatic differentiation of human embryonic stem cells (hESCs). Methods hESC-derived HLCs were generated through multistage differentiation in two-dimensional (2D) and three-dimensional (3D) cultures, incorporating either type I collagen or Matrigel during hepatic specification and maturation. The resulting HLCs was characterized for their gene expression and functionality using various molecular and cellular techniques. Results Our results showed that HLCs cultured with collagen exhibited a significant increase in albumin and alpha-1 anti-trypsin expression with reduced AFP compared to HLCs cultured with Matrigel. They also secreted more urea than Matrigel cultures. However, these HLCs exhibited lower CYP3A4 activity and glycogen storage than those cultured with Matrigel. These functional differences in HLCs between collagen and Matrigel cultures closely resembled the hepatocytes of periportal and pericentral zones, respectively. Conclusion Our study demonstrates that Matrigel and collagen have differential effects on the differentiation and functionality of HLCs, which resemble, to an extent, hepatic zonation in the liver lobules. Our finding has an important impact on the generation of hPSC-HLCs for biomedical and medical applications. |
| first_indexed | 2024-03-11T12:43:37Z |
| format | Article |
| id | doaj.art-49634cbe55714741bb334873f0bdec14 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-03-11T12:43:37Z |
| publishDate | 2023-11-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-49634cbe55714741bb334873f0bdec142023-11-05T12:09:27ZengBMCStem Cell Research & Therapy1757-65122023-11-0114111110.1186/s13287-023-03542-xExtracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cellsFaiza Farhan0Manjari Trivedi1Priscilla Di Wu2Wei Cui3Institute of Reproductive and Developmental Biology, Faculty of Medicine, Imperial College LondonInstitute of Reproductive and Developmental Biology, Faculty of Medicine, Imperial College LondonInstitute of Reproductive and Developmental Biology, Faculty of Medicine, Imperial College LondonInstitute of Reproductive and Developmental Biology, Faculty of Medicine, Imperial College LondonAbstract Background Human pluripotent stem cell (hPSC)-derived hepatocyte-like cells (HLCs) can provide a valuable in vitro model for disease modelling and drug development. However, generating HLCs with characteristics comparable to hepatocytes in vivo is challenging. Extracellular matrix (ECM) plays an important role in supporting liver development and hepatocyte functions, but their impact on hepatocyte differentiation and maturation during hPSC differentiation remains unclear. Here, we investigate the effects of two ECM components—Matrigel and type I collagen on hepatic differentiation of human embryonic stem cells (hESCs). Methods hESC-derived HLCs were generated through multistage differentiation in two-dimensional (2D) and three-dimensional (3D) cultures, incorporating either type I collagen or Matrigel during hepatic specification and maturation. The resulting HLCs was characterized for their gene expression and functionality using various molecular and cellular techniques. Results Our results showed that HLCs cultured with collagen exhibited a significant increase in albumin and alpha-1 anti-trypsin expression with reduced AFP compared to HLCs cultured with Matrigel. They also secreted more urea than Matrigel cultures. However, these HLCs exhibited lower CYP3A4 activity and glycogen storage than those cultured with Matrigel. These functional differences in HLCs between collagen and Matrigel cultures closely resembled the hepatocytes of periportal and pericentral zones, respectively. Conclusion Our study demonstrates that Matrigel and collagen have differential effects on the differentiation and functionality of HLCs, which resemble, to an extent, hepatic zonation in the liver lobules. Our finding has an important impact on the generation of hPSC-HLCs for biomedical and medical applications.https://doi.org/10.1186/s13287-023-03542-xExtracellular matrix (ECM)HepatocytehESC differentiationHepatic zonationMatrigelCollagen |
| spellingShingle | Faiza Farhan Manjari Trivedi Priscilla Di Wu Wei Cui Extracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cells Stem Cell Research & Therapy Extracellular matrix (ECM) Hepatocyte hESC differentiation Hepatic zonation Matrigel Collagen |
| title | Extracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cells |
| title_full | Extracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cells |
| title_fullStr | Extracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cells |
| title_full_unstemmed | Extracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cells |
| title_short | Extracellular matrix modulates the spatial hepatic features in hepatocyte-like cells derived from human embryonic stem cells |
| title_sort | extracellular matrix modulates the spatial hepatic features in hepatocyte like cells derived from human embryonic stem cells |
| topic | Extracellular matrix (ECM) Hepatocyte hESC differentiation Hepatic zonation Matrigel Collagen |
| url | https://doi.org/10.1186/s13287-023-03542-x |
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