Abstract
In response to liver injury, hepatic cells, especially hepatocytes, can rapidly proliferate to repair liver damage. Additionally, it was shown that under certain circumstances liver resident cells with progenitor capabilities are involved in liver cell proliferation and differentiation. These hepatic progenitor cells (HPCs), known as oval cells in rodents, are derived from the canals of Hering, which are located in the periportal region of the liver. Regarding to different cell niches, which were defined for human HPCs, several markers have been used to identify these cells such as CD34, c-kit, OV-6, and Thy-1 (CD90). The latter was shown to be expressed on HPCs in human liver tissue with histological signs of regeneration. In this chapter we describe a detailed method for the isolation of Thy-1 positive cells from human resected liver tissue. Based on a procedure for isolating primary human hepatocytes and non-parenchymal cells (NPCs) we expanded this protocol to additional enzymatic dissociation, filtration, and centrifugation steps. This results in a bile duct cell enriched fraction of NPCs from which Thy-1 (CD90) positive cells were purified by Thy-1 positivity selection using MACS technique. Bipotential progenitor cells from human liver resections can be isolated using Thy-1 and was shown to be a suitable tool for the enrichment of liver resident progenitor cells for xenotransplantation.
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References
Michalopoulos GK, DeFrances MC (1997) Liver regeneration. Science 276:60–66
Michalopoulos GK (2010) Liver regeneration after partial hepatectomy: critical analysis of mechanistic dilemmas. Am J Pathol 176:2–13
Fausto N (2004) Liver regeneration and repair: hepatocytes, progenitor cells, and stem cells. Hepatology 39:1477–1487
Roskams TA, Theise ND, Balabaud C, Bhagat G, Bhathal PS, Bioulac-Sage P et al (2004) Nomenclature of the finer branches of the biliary tree: canals, ductules, and ductular reactions in human livers. Hepatology 39:1739–1745
Fausto N, Campbell JS (2003) The role of hepatocytes and oval cells in liver regeneration and repopulation. Mech Dev 120:117–130
Walkup MH, Gerber DA (2006) Hepatic stem cells: in search of. Stem Cells 24:1833–1840
Terrace JD, Currie IS, Hay DC, Masson NM, Anderson RA, Forbes SJ et al (2007) Progenitor cell characterization and location in the developing human liver. Stem Cells Dev 16:771–778
Weiss TS, Lichtenauer M, Kirchner S, Stock P, Aurich H, Christ B et al (2008) Hepatic progenitor cells from adult human livers for cell transplantation. Gut 57:1129–1138
Tolosa L, Bonora-Centelles A, Teresa DM, Pareja E, Negro A, Lopez S et al (2011) Steatotic liver: a suitable source for the isolation of hepatic progenitor cells. Liver Int 31:1231–1238
Masson NM, Currie IS, Terrace JD, Garden OJ, Parks RW, Ross JA (2006) Hepatic progenitor cells in human fetal liver express the oval cell marker Thy-1. Am J Physiol Gastrointest Liver Physiol 291:G45–G54
Ono Y, Kawachi S, Hayashida T, Wakui M, Tanabe M, Itano O et al (2011) The influence of donor age on liver regeneration and hepatic progenitor cell populations. Surgery 150:154–161
Damm G, Pfeiffer E, Burkhardt B, Vermehren J, Nussler AK, Weiss TS (2013) Human parenchymal and non-parenchymal liver cell isolation, culture and characterization. Hepatol Int 7:951–958
Godoy P, Hewitt NJ, Albrecht U, Andersen ME, Ansari N, Bhattacharya S et al (2013) Recent advances in 2D and 3D in vitro systems using primary hepatocytes, alternative hepatocyte sources and non-parenchymal liver cells and their use in investigating mechanisms of hepatotoxicity, cell signaling and ADME. Arch Toxicol 87:1315–1530
Weiss TS, Jahn B, Cetto M, Jauch KW, Thasler WE (2002) Collagen sandwich culture affects intracellular polyamine levels of human hepatocytes. Cell Prolif 35:257–267
Dayoub R, Vogel A, Schuett J, Lupke M, Spieker SM, Kettern N et al (2013) Nrf2 activates augmenter of liver regeneration (ALR) via antioxidant response element and links oxidative stress to liver regeneration. Mol Med 19:237–244
Acknowledgement
The authors would like to thank Susanne Heyn for her excellent technical assistance. This study was supported by the German Federal Ministry of Education and Research (Virtual Liver Network Grants 0315753 to T.S.W.)
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Weiss, T.S., Dayoub, R. (2017). Thy-1 (CD90)-Positive Hepatic Progenitor Cells, Hepatoctyes, and Non-parenchymal Liver Cells Isolated from Human Livers. In: Stock, P., Christ, B. (eds) Hepatocyte Transplantation. Methods in Molecular Biology, vol 1506. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6506-9_5
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DOI: https://doi.org/10.1007/978-1-4939-6506-9_5
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Publisher Name: Humana Press, New York, NY
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