Wang Gaoxiong, Zheng Youshi, Wang Yingchao, Cai Zhixiong, Liao Naishun, Liu Jingfeng, Zhang Wenmin
Department of Hepatobiliary Surgery, The Second Affiliated Hospital of Fujian Medical University, Quanzhou, 362000, People's Republic of China.
The United Innovation of Mengchao Hepatobiliary Technology Key Laboratory of Fujian Province, Mengchao Hepatobiliary Hospital of Fujian Medical University, Fuzhou, 350025, People's Republic of China.
Cytotechnology. 2018 Aug;70(4):1279-1290. doi: 10.1007/s10616-018-0219-3. Epub 2018 Apr 19.
Although hepatocyte transplantation and bioartificial liver support system provide new promising opportunities for those patients waiting for liver transplantation, hepatocytes are easily losing liver-specific functions by using the common in vitro cultured methods. The co-culture strategies with mimicking the in vivo microenvironment would facilitate the maintenance of liver-specific functions of hepatocytes. Considering that hepatocytes and endothelial cells (ECs) account for 80-90% of total cell populations in the liver, hepatocytes and ECs were directly co-cultured with hepatic stellate cells (HSCs) or adipose tissue-derived stem cells (ADSCs) at a ratio of 700:150:3 or 14:3:3 in the present study, and the liver-specific functions were carefully analyzed. Our results showed that the two co-culture systems presented the enhanced liver-specific functions through promoting secretion of urea and ALB and increasing the expressions of ALB, CYP3A4 and HNF4α, and the vessel-like structure in the co-culture system consisted of hepatocytes, ECs and ADSCs. Hence, our results suggested that the directly co-culture of hepatocytes and ECs with HSCs or ADSCs could significantly improve liver-specific functions of hepatocytes, and the co-culture system could further promote angiogenesis of ECs at a later stage. Therefore, this study provides potential interesting in vitro strategies for enhancing liver-specific functions of hepatocytes.
尽管肝细胞移植和生物人工肝支持系统为等待肝移植的患者提供了新的、有前景的机会，但采用常见的体外培养方法时，肝细胞很容易丧失肝脏特异性功能。模拟体内微环境的共培养策略将有助于维持肝细胞的肝脏特异性功能。鉴于肝细胞和内皮细胞（ECs）占肝脏细胞总数的80-90%，在本研究中，肝细胞和ECs分别与肝星状细胞（HSCs）或脂肪组织来源的干细胞（ADSCs）以700:150:3或14:3:3的比例直接共培养，并对肝脏特异性功能进行了仔细分析。我们的结果表明，这两种共培养系统通过促进尿素和白蛋白的分泌以及增加白蛋白、细胞色素P450 3A4（CYP3A4）和肝细胞核因子4α（HNF4α）的表达，呈现出增强的肝脏特异性功能，并且共培养系统中的血管样结构由肝细胞、ECs和ADSCs组成。因此，我们的结果表明，肝细胞和ECs与HSCs或ADSCs直接共培养可显著改善肝细胞的肝脏特异性功能，并且该共培养系统在后期可进一步促进ECs的血管生成。因此，本研究为增强肝细胞的肝脏特异性功能提供了潜在的、有趣的体外策略。
