Coculture With Mesenchymal Stem Cells Results in Improved Viability and Function of Human Hepatocytes
Coculture With Mesenchymal Stem Cells Results in Improved Viability and Function of Human Hepatocytes
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DOI：
10.3727/096368913x674080
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发表时间：
2015-01-01
影响因子：
3.3
通讯作者：
Dhawan, Anil
中科院分区：
文献类型：
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作者：
Fitzpatrick, Emer;Wu, Yue;Dhawan, Anil
肝细胞移植正成为急性肝功能衰竭的公认治疗方法，无论是作为肝脏再生还是器官移植的桥梁。肝细胞代替衰竭的器官提供肝功能。维持移植肝细胞足够的活力和功能是一个问题。近年来，间充质干细胞（MSC）为肝细胞提供结构和营养支持引起了广泛的兴趣，但目前很少有研究使用原代人肝细胞。本研究的目的是研究人骨髓间充质干细胞与冻存的人肝细胞共培养是否可以改善其功能和活力，从而具有肝病细胞治疗的潜力。从人脐带或脂肪组织中分离MSC。从不适合移植的供体器官中分离肝细胞。骨髓间充质干细胞和肝细胞直接和间接接触共培养。来自共培养的MSC和肝细胞的条件培养基（CM）也用于肝细胞。比较了试验组和对照组之间的活力和肝脏特异性功能。直接与MSC共培养的人肝细胞显示从培养的第5天至第25天白蛋白的产生得到改善。这种影响在第15天最为突出。同样，尿素产量在共培养中从第5天到第25天得到改善。间接共培养证明，与肝细胞单一培养（940 ng/ml）相比，第4天的白蛋白产量（1，107 ng/ml）有所改善。CM中的肝细胞显示功能无显著改善。共培养肝细胞的活力优于单培养细胞，提高了16%。因此，人肝细胞与MSC的共培养证明了改善的功能和活力。这种效应主要见于直接共培养，但也见于间接培养和CM。这样的共培养条件可以传达肝细胞存活和细胞移植功能的主要优势。
Hepatocyte transplantation is becoming an accepted therapy for acute liver failure, either as a bridge to liver regeneration or to organ transplantation. Hepatocytes provide liver function in place of the failing organ. The maintenance of sufficient viability and function of the transplanted hepatocytes is a concern. There is a lot of recent interest in mesenchymal stem cells (MSCs) for the provision of structural and trophic support to hepatocytes, but few studies currently use primary human hepatocytes. The aim of this study was to investigate if coculture of human MSCs with cryopreserved human hepatocytes may improve their function and viability, thus with potential for cellular therapy of liver disease. MSCs were isolated from human umbilical cord or adipose tissue. Hepatocytes were isolated from donor organs unsuitable for transplantation. MSCs and hepatocytes were cocultured in both direct and indirect contact. Conditioned medium (CM) from cocultured MSCs and hepatocytes was also used on hepatocytes. Viability and liver-specific function were compared between test and controls. Human hepatocytes that were cocultured directly with MSCs demonstrated improved production of albumin from day 5 to day 25 of culture. This effect was most prominent at day 15. Likewise, urea production was improved in coculture from day 5 to 25. Indirect coculture demonstrated improved albumin production by day 4 (1,107 ng/ml) versus hepatocyte monoculture (940 ng/ml). Hepatocytes in CM demonstrated a nonsignificant improvement in function. The viability of cocultured hepatocytes was superior to that of monocultured cells with up to a 16% improvement. Thus, coculture of human hepatocytes with MSCs demonstrates both improved function and viability. The effect is seen mainly with direct coculture but can also be seen in indirect culture and with CM. Such coculture conditions may convey major advantages in hepatocyte survival and function for cell transplantation.