Culture of newborn monkey liver epithelial progenitor cells in chemical defined serum-free medium.
Studies with hepatic progenitor cells from non-human primates would allow better understanding of their human counterparts. In this study, rhesus monkey liver epithelial progenitor cells (mLEPCs) were derived from a small piece of newborn livers in chemical defined serum-free medium. Digested hepatic cells were treated in Ca(2+)-containing medium to form cell aggregates. Two types of cell aggregates were generated: elongated spindle cells and polygonal epithelial cells. Elongated spindle cells were expressed as vimentin and brachyury, and they were disappeared within 5 d in our cultures. The remaining type 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 alpha-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 alpha-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. Thus, mLEPCs with bipotency were derived from newborn monkey liver and may serve as a preclinical model for assessment of cell therapy in humans.
Ji
Shaohui
S
Kunming Primate Research Center, and Kunming Institute of Zoology, Chinese Academy of Sciences, 32# Jiaochang Donglu, Kunming, Yunnan, 650223, China.
Jin
Lifang
L
Guo
Xiangyu
X
Ji
Weizhi
W
D000818:Animals; D000831:Animals, Newborn; D015820:Cadherins / Q000032:analysis; D002449:Cell Aggregation; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D016895:Culture Media, Serum-Free; D003907:Dexamethasone / Q000494:pharmacology; D004847:Epithelial Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D039503:Integrin alpha6 / Q000032:analysis; D007381:Intermediate Filament Proteins / Q000032:analysis; D053531:Keratins, Type II / Q000032:analysis; D008099:Liver / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D008253:Macaca mulatta; D051379:Mice; D009419:Nerve Tissue Proteins / Q000032:analysis; D064231:Nestin; D053683:Oncostatin M / Q000494:pharmacology; D051381:Rats; D013234:Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D000515:alpha 1-Antitrypsin / Q000032:analysis
D015820:Cadherins; D016895:Culture Media, Serum-Free; D039503:Integrin alpha6; D007381:Intermediate Filament Proteins; D053531:Keratins, Type II; C577484:NES protein, human; D009419:Nerve Tissue Proteins; C577485:Nes protein, mouse; C577486:Nes protein, rat; D064231:Nestin; D000515:alpha 1-Antitrypsin; D053683:Oncostatin M; D003907:Dexamethasone