Faculty of Dentistry, University of Toronto, 124 Edward Street, Toronto, ON M5G 1G6, Canada.
Biochem Cell Biol. 2013 Jun;91(3):140-7. doi: 10.1139/bcb-2012-0079. Epub 2012 Nov 28.
Hepatocyte culture is a useful tool for the study of their biology and the development of bioartificial livers. However, many challenges have to be overcome since hepatocytes rapidly lose their normal phenotype in vitro. We have recently demonstrated that human umbilical cord perivascular cells (HUCPVCs) are able to provide support to hepatocytes. In the present study we go further into exploring the effects that HUCPVCs have in the functional polarization, and both the internal and external organization, of hepatocytes. Also, we investigate HUCPVC-hepatocyte crosstalk by tracking both the effects of HUCPVCs on hepatocyte transcription factors and those of hepatocytes on the expression of hepatotrophic factors in HUCPVCs. Our results show that HUCPVCs maintain the functional polarity of hepatocytes ex vivo, as judged by the secretion of fluorescein into bile canaliculi, for at least 40 days. Transmission electron microscopy revealed that hepatocytes in coculture organize in an organoid-like structure embedded in extracellular matrix surrounded by HUCPVCs. In coculture, hepatocytes displayed a higher expression of C/EBPα, implicated in maintenance of the mature hepatocyte phenotype, and HUCPVCs upregulated hepatocyte growth factor and Jagged1 indicating that these genes may play important roles in HUCPVC-hepatocyte interactions.
肝细胞培养是研究其生物学特性和生物人工肝脏发展的有用工具。然而，由于肝细胞在体外迅速失去其正常表型，因此需要克服许多挑战。我们最近证明，人脐带血周细胞（HUCPVCs）能够为肝细胞提供支持。在本研究中，我们进一步探讨了 HUCPVCs 对肝细胞功能极化以及内部和外部组织的影响。此外，我们还通过跟踪 HUCPVCs 对肝细胞转录因子的影响以及肝细胞对 HUCPVCs 中肝营养因子表达的影响，研究了 HUCPVC-肝细胞的串扰。我们的结果表明，HUCPVCs 至少能在体外表征肝细胞的功能极性，通过将荧光素分泌到胆小管来判断，至少能持续 40 天。透射电子显微镜显示，共培养中的肝细胞在由 HUCPVCs 包围的细胞外基质中组织成类器官样结构。在共培养中，肝细胞表达更高水平的 C/EBPα，这与维持成熟肝细胞表型有关，HUCPVCs 上调了肝细胞生长因子和 Jagged1，表明这些基因可能在 HUCPVC-肝细胞相互作用中发挥重要作用。
