Abstract
Bipotent liver progenitor cells (LPCs) are promising cell sources for cell transplantation therapy in hepatic disorders as well as biliary dysfunctions. Using a cocktail of small molecules, we recently reported a novel approach to generate bipotent LPCs, named chemically induced liver progenitors (CLiPs), from adult rat hepatocytes. In this chapter, we describe a detailed protocol for the induction of rat CLiPs. We first describe the method to isolate primary rat hepatocytes and then describe how to induce CLiPs from the hepatocytes. In addition, we describe methods to induce the generated CLiPs to differentiate into hepatocytes and biliary epithelial cells.
Key words
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Woo DH, Kim SK, Lim HJ et al (2012) Direct and indirect contribution of human embryonic stem cell-derived hepatocyte-like cells to liver repair in mice. Gastroenterology 142:602–611. https://doi.org/10.1053/j.gastro.2011.11.0301
Oertel M, Menthena A, Chen Y-Q et al (2008) Purification of fetal liver stem/progenitor cells containing all the repopulation potential for normal adult rat liver. Gastroenterology 134:823–832. https://doi.org/10.1053/j.gastro.2008.01.007
Liu H, Kim Y, Sharkis S et al (2011) In vivo liver regeneration potential of human induced pluripotent stem cells from diverse origins. Sci Transl Med 3:82ra39. https://doi.org/10.1126/scitranslmed.3002376
Zhu S, Rezvani M, Harbell J et al (2014) Mouse liver repopulation with hepatocytes generated from human fibroblasts. Nature 508:93–97. https://doi.org/10.1038/nature13020
Sekiya S, Suzuki A (2011) Direct conversion of mouse fibroblasts to hepatocyte-like cells by defined factors. Nature 475:390–393. https://doi.org/10.1038/nature10263
Huang P, He Z, Ji S et al (2011) Induction of functional hepatocyte-like cells from mouse fibroblasts by defined factors. Nature 475:386–389. https://doi.org/10.1038/nature10116
Huch M, Dorrell C, Boj SF et al (2013) In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature 494:247–250. https://doi.org/10.1038/nature11826
Lu W-Y, Bird TG, Boulter L et al (2015) Hepatic progenitor cells of biliary origin with liver repopulation capacity. Nat Cell Biol 17:971–983. https://doi.org/10.1038/ncb3203
Katsuda T, Kawamata M, Hagiwara K et al (2017) Conversion of terminally committed hepatocytes to culturable bipotent progenitor cells with regenerative capacity. Cell Stem Cell 20:41–55. https://doi.org/10.1016/j.stem.2016.10.007
Seglen PO (1976) Preparation of isolated rat liver cells. Methods Cell Biol 13:29–83. https://doi.org/10.1016/S0091-679X(08)61797-5
Loyer P, Cariou S, Glaise D et al (1996) Growth factor dependence of progression through G1 and S phases of adult rat hepatocytes in vitro. Evidence of a mitogen restriction point in mid-late G1. J Biol Chem 271:11484–11492
Frémin C, Bessard A, Ezan F et al (2009) Multiple division cycles and long-term survival of hepatocytes are distinctly regulated by extracellular signal-regulated kinases ERK1 and ERK2. Hepatology 49:930–939. https://doi.org/10.1002/hep.22730
Kamiya A, Kojima N, Kinoshita T et al (2002) Maturation of fetal hepatocytes in vitro by extracellular matrices and oncostatin M: induction of tryptophan oxygenase. Hepatology 35:1351–1359. https://doi.org/10.1053/jhep.2002.33331
Acknowledgments
This work was supported in part by Grant-in-Aid for Young Scientists B (16 K16643) and the Research Program on Hepatitis (16fk0310505h0005) from the Japan Agency for Medical Research and Development (AMED).
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Katsuda, T., Ochiya, T. (2019). Chemically Induced Liver Progenitors (CLiPs): A Novel Cell Source for Hepatocytes and Biliary Epithelial Cells. In: Tanimizu, N. (eds) Hepatic Stem Cells . Methods in Molecular Biology, vol 1905. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8961-4_11
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8961-4_11
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8960-7
Online ISBN: 978-1-4939-8961-4
eBook Packages: Springer Protocols