Gu Jinyang, Shi Xiaolei, Zhang Yue, Ding Yitao
Department of Hepatobiliary Surgery, DrumTower Clinical Medical College of Nanjing Medical University, Nanjing, China.
J Cell Physiol. 2009 Apr;219(1):100-8. doi: 10.1002/jcp.21651.
Temporary replacement of specific liver functions with extracorporeal bioartificial liver has been hampered by rapid de-differentiation of porcine hepatocytes in vitro. Co-cultivation of hepatocytes with non-parenchymal cells may be beneficial for optimizing cell functions via mimicry of physiological microenvironment consisting of endogenous matrix proteins. However, the underlying mechanisms remain to be elucidated. A randomly distributed co-culture system composed of porcine hepatocytes and bone marrow mesenchymal stem cells was generated, and the morphological and functional changes of varying degrees of heterotypic interactions were characterized. Furthermore, contributions of extracellular matrix within this co-culture were evaluated. A rapid attachment and self-organization of three-dimensional hepatocyte spheroids were encouraged. Studies on hepatocyte viability showed a metabolically active, viable cell population in all co-culture configurations with occurrence of few dead cells. The maximal induction of albumin production, urea synthesis, and cytochrome P4503A1 activities was achieved at seeding ratio of 2:1. Immunocytochemical detection of various extracellular matrix confirmed that a high level of matrix proteins synthesis within distinct cells was involved in hepatocyte homeostasis. These results demonstrate for the first time that cell-matrix has synergic effects on the preservation of hepatic morphology and functionality in the co-culture of porcine hepatocytes with mesenchymal stem cells in vitro, which could represent a promising tool for tissue engineering, cell biology, and bioartificial liver devices.
体外猪肝细胞的快速去分化阻碍了用体外生物人工肝暂时替代特定肝功能。肝细胞与非实质细胞共培养可能有助于通过模拟由内源性基质蛋白组成的生理微环境来优化细胞功能。然而，其潜在机制仍有待阐明。构建了一个由猪肝细胞和骨髓间充质干细胞组成的随机分布共培养系统，并对不同程度异型相互作用的形态和功能变化进行了表征。此外，还评估了该共培养体系中细胞外基质的作用。促进了三维肝细胞球的快速附着和自我组织。肝细胞活力研究表明，在所有共培养组合中均存在代谢活跃、存活的细胞群体，死细胞数量很少。在接种比例为2:1时，白蛋白产生、尿素合成和细胞色素P4503A1活性达到最大诱导水平。对各种细胞外基质的免疫细胞化学检测证实，不同细胞内高水平的基质蛋白合成参与了肝细胞内稳态。这些结果首次表明，在体外猪肝细胞与间充质干细胞共培养中，细胞-基质对肝脏形态和功能的维持具有协同作用，这可能是组织工程、细胞生物学和生物人工肝装置的一个有前途的工具。
