Growth and differentiation of colony-forming human hepatocytes in vitro
Growth and differentiation of colony-forming human hepatocytes in vitro
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DOI：
10.1016/j.jhep.2005.10.028
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发表时间：
2006-04-01
影响因子：
25.7
通讯作者：
Yoshizato, K
中科院分区：
文献类型：
--
作者：
Yamasaki, C;Tateno, C;Yoshizato, K
背景/目的：大鼠肝实质细胞(PHS)含有一小部分集落形成的实质肝细胞(CF-PHS)。我们的目的是证明人CFPHS的存在，并从生长和分化能力方面对其进行表征。方法：将人PHS与瑞士3T3细胞在含有人血清、EGF、尼康替胺和抗坏血酸2-磷酸的培养液中共同培养。在Matrigel Matrix凝胶上培养肝细胞以检测分化潜能。结果：PHS几乎没有形成集落，集落形成率仅为0.01-0.09%。CF-PHS可传代至7代。融合前和融合早期呈肝上皮细胞样形态，白蛋白(ALB)、细胞角蛋白(CK)7、8、18和19呈免疫细胞化学阳性；融合时呈典型的分化肝细胞样形态，α(1)-抗胰蛋白酶阳性，凝集区CK7和19阴性。融合晚期的Cf-PHS低水平表达ALB、HNF4和细胞色素P450亚型的mRNA。然而，当这些细胞在Matrigel上培养时，它们的表达水平与原始PH相当。结论：我们得出结论：人类肝脏含有高度复制的肝祖细胞样细胞，作为一个保持正常分化潜能的微小群体。(C)2005年欧洲肝脏研究协会。爱思唯尔出版，版权所有。
Background/Aims: Parenchymal hepatocytes (PHs) of rat contain colony-forming parenchymal hepatocytes (CF-PHs) as a small fraction. We aimed to demonstrate the presence of CF-PHs in humans and characterize them with respect to growth and differentiation potential.Methods: Human PHs were co-cultured with Swiss 3T3 cells in the medium containing human serum, EGF, nicontinamide, and ascorbic acid 2-phosphate. To examine differentiation potential hepatocytes were cultured on gels of Matrigel Matrix.Results: Few PHs formed colonies, the colony-forming efficiency being as low as 0.01-0.09%. The CF-PHs could be subcultured up to 7 passages. They showed a liver epithelial cell-like morphology, and immunocytochemically positive for albumin (ALB), cytokeratin (CK) 7, 8, 18, and 19 in a pre- and early phase-confluence, whereas they showed a typical differentiated hepatocyte-like morphology, and positive for alpha(1)-antitrypsin, but negative for CK7 and 19 in condensed regions at confluence. The CF-PHs at late confluence expressed mRNAs of ALB, HNF4, and isoforms of cytochrome P450 at low levels. However, when cultured on Matrigel, these cells expressed them at high levels comparable to those of original PHs.Conclusions: We concluded that the human liver contains highly replicative hepatic progenitor-like cells as a minute population that retain a normal differentiation potential. (c) 2005 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.