Laboratory of Stem Cells, Institute of Cell Biology, College of Life Sciences, Zhejiang University, Hangzhou, Zhejiang, People's Republic of China.
J Tissue Eng Regen Med. 2012 Jan;6(1):29-39. doi: 10.1002/term.393. Epub 2011 Mar 10.
Bone marrow mesenchymal stem cells (MSCs) are promising candidates for cell therapy and tissue engineering. We used mesenchymal stem cells from human bone marrow (hMSCs) as the seeding cells to investigate the potential of hepatocytic differentiation of hMSCs in porous polylactic glycolic acid (PLGA) scaffolds under perfusion induction. hMSCs were seeded and proliferated in PLGA scaffolds, and then induced into hepatocyte-like cells with hepatogenic medium in perfusion and static cultures. The results showed that hMSCs could be induced into hepatocyte-like cells in PLGA scaffolds with hepatogenic medium in both static and perfusion induction systems. However, perfusion induction was more effective for cellularity in PLGA scaffolds than in static induction. Cells in the scaffold induced by the hepatogenic medium expressed hepatocyte-specific genes cytokeratin 19 (CK19), α-fetoprotein (αFP), cytokeratin 18 (CK18), albumin and cytochrome P4503A4 (CYP3A4) in a time-dependent manner. Induced cells stained positive for αFP and albumin. Induced cells also acquired the functional characteristics of hepatocytes, i.e. secretion of urea and albumin. In a comparison of survival and hepatogenic differentiation of hMSCs between perfusion and static induction, perfusion induction increased the survival and the uniform distribution of induced cells in scaffolds, which resulted in a higher efficiency of hepatogenesis in the PLGA construct with hMSCs. The oscillatory perfusion induction system combined with the hepatogenic medium should be a valuable and convenient tool for in vitro hepatic tissue engineering using hMSCs.
骨髓间充质干细胞（MSCs）是细胞治疗和组织工程的有前途的候选者。我们使用人骨髓间充质干细胞（hMSCs）作为种子细胞，在灌注诱导下，研究多孔聚乳酸-乙醇酸（PLGA）支架中 hMSCs 向肝样细胞分化的潜力。将 hMSCs 接种并在 PLGA 支架中增殖，然后用肝生成培养基在灌注和静态培养中诱导为肝样细胞。结果表明，hMSCs 可以在肝生成培养基的 PLGA 支架中被诱导为肝样细胞，无论是在静态还是灌注诱导系统中。然而，与静态诱导相比，灌注诱导更有利于细胞在 PLGA 支架中的增殖。用肝生成培养基诱导的支架中的细胞以时间依赖性方式表达肝特异性基因细胞角蛋白 19（CK19）、甲胎蛋白（αFP）、细胞角蛋白 18（CK18）、白蛋白和细胞色素 P4503A4（CYP3A4）。诱导的细胞对 αFP 和白蛋白呈阳性染色。诱导的细胞还获得了肝细胞的功能特性，即尿素和白蛋白的分泌。在灌注和静态诱导对 hMSCs 的存活和肝向分化的比较中，灌注诱导增加了诱导细胞在支架中的存活和均匀分布，从而使 hMSCs 在 PLGA 构建体中的肝发生效率更高。振荡灌注诱导系统与肝生成培养基相结合，应该是使用 hMSCs 进行体外肝组织工程的一种有价值且方便的工具。
