Culture of Newborn Rhesus Monkey Liver Epithelial Progenitor Cells in Chemical Defined Serum-free Medium

doi:10.3969/j.issn.1674-5817.2011.01.005

Abstract

Abstract: Objective to establish a chemical defined serum-free culture system for rhesus monkey hepatic progenitor cells. Methods a rhesus monkey liver polygon cell colony was derived from a small piece of newborn livers using chemical defined serum-free medium. Follow by isolated and cultured in different extracellular matrix. And the identification of the cells phenotype and dual-differentiation potential for functional hepatocytes and biliary duct cells in vitro were carry out using immunofluorescence cell staining, Periodic Acid-Shiff Staining, RT-PCR, indocyanine green (ICG) uptake staining method. Result Two types of cell colony were generated: elongated spindle cells and polygonal epithelial cells. Elongated spindle cells were expressed as vimentin and brachyury, and they were disappeared within 5 days in our cultures. The remaining consisted of small polygonal epithelial cells that expressed cytokeratin 7 (CK7), CK8, CK18, nestin, CD49f and E-cad, the markers of hepatic stem cells, but were negative for 〈-fetoprotein, albumin and CK19. They can proliferate and be passaged, if on laminin or rat tail collagen gel, to initiate colonies. When cultured with dexamethasone and oncostatin M, the expression of mature hepatocyte markers, such as 〈-1-antitrypsin, intracytoplasmic glycogen storage, indocyanine green uptake and lipid droplet generation, were induced in differentiated cells. If transferred onto mouse embryonic fibroblasts feeders, they gave rise to CK19 positive cholangiocytes with formation of doughnut-like structure. Conclusion we isolated a polygon epithelial cell with characteristics of live progenitor cells from newborn rhesus monkey live tissue using a chemical defined serum-free culture system, and. may serve as an in vitro model of primate liver development and may facilitate further preclinical research.

Key words: Hepatic progenitor cells, Dual-differentiation potential, Serum free, Extracellullar matrix

SUN Ai-jing, Huang Di-lai, RU Guo-mei, JIN Li-fang. Culture of Newborn Rhesus Monkey Liver Epithelial Progenitor Cells in Chemical Defined Serum-free Medium[J]. Laboratory Animal and Comparative Medicine, 2011, 31(1): 20-27.

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