ABSTRACT
Differentiation of stem cells to hepatocytes provides an unlimited supply of human hepatocytes and therefore has been vigorously studied. However, to date, the stem cell-derived hepatocytes were suggested to be of immature features. To obtain matured hepatocytes from stem cells, we tested the effect of culturing human-induced pluripotent stem (hiPS) cell-derived endoderm cells on collagen vitrigel membrane and compared with our previous reported nanofiber matrix. We cultured hiPS cell-derived endoderm cells on a collagen vitrigel membrane and examined the expression profiles, and tested the activity of metabolic enzymes. Gene expression profile analysis of hepatocytic differentiation markers revealed that upon culture on collagen vitrigel membrane, immature markers of AFP decreased, with a concomitant increase in the expression of mature hepatocyte transcription factors and mature hepatocyte markers such as ALB, ASGR1. Mature markers involved in liver functions, such as transporters, cytochrome P450 enzymes and phase II metabolic enzymes were also upregulated. We observed the upregulation of the liver markers for at least 2 weeks. Gene array profiling analysis revealed that hiPS cell-derived hepatocyte-like cells (hiPS-hep) resemble those of the primary hepatocytes. Functions of the CYP enzyme activities were tested in multi-institution and all revealed high CYP1A, CYP2C19, CYP2D6, CYP3A activity, which could be maintained for at least 2 weeks in culture. Taken together, the present approach identified that collagen vitrigel membrane provides a suitable environment for the generation of hepatocytes from hiPS cells that resemble many characteristics of primary human hepatocytes.
Footnotes
Competing interests
H.Y. and N.T. are employees in Kanto Chemical Co. Inc., which supplies collagen vitrigel membrane products.
Author contributions
Conceptualization: S.N., N.S., S.K.; Methodology: S.N., H.Y., N.S., S.K.; Validation: S.N., I.S., T.S., H.Y., N.T., T.I., T.N., J.K., S.W., T.M., E.H., S.I., N.S., S.K.; Formal analysis: S.N., H.Y., T.I., T.N., J.K., S.W., T.M., E.H., N.S., S.K.; Investigation: S.N., I.S., T.S., H.Y., N.T., T.I., T.N., J.K., S.W., T.M., E.H., N.S., S.K.; Resources: N.T., N.S., S.K.; Data curation: H.Y., T.I., T.N., J.K., S.W., T.M., E.H., S.I., N.S., S.K.; Writing - original draft: S.K.; Writing - review & editing: I.S., T.S., H.Y., N.T., T.I., T.N., J.K., S.W., T.M., E.H., S.I., N.S., S.K.; Visualization: S.K.; Supervision: N.S., S.K.; Project administration: N.S., S.K.; Funding acquisition: S.K.
Funding
This work was supported by a grant from Japan Agency for Medical Research and Development (AMED) (to S.K., Grant no. 18bk0104075h0102).
Data availability
Data can be found at the Dryad Digital Repository: https://datadryad.org/resource/doi:10.5061/dryad.5f3s52r/1.
Supplementary information
Supplementary information available online at http://bio.biologists.org/lookup/doi/10.1242/bio.042192.supplemental
- Received February 1, 2019.
- Accepted May 19, 2019.
- © 2019. Published by The Company of Biologists Ltd
This is an Open Access article distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.
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