Rotating Microgravity-Bioreactor Cultivation Enhances the Hepatic Differentiation of Mouse Embryonic Stem Cells on Biodegradable Polymer Scaffolds
Rotating Microgravity-Bioreactor Cultivation Enhances the Hepatic Differentiation of Mouse Embryonic Stem Cells on Biodegradable Polymer Scaffolds
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DOI：
10.1089/ten.tea.2012.0097
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发表时间：
2012-11-01
影响因子：
4.1
通讯作者：
Geng, Yongjian
中科院分区：
文献类型：
--
作者：
Wang, Yingjie;Zhang, Yunping;Geng, Yongjian
胚胎干细胞是一种多能细胞，能够分化所有的体细胞系，包括肝组织中的体细胞系。我们描述了使用可生物降解的聚合物支架和允许模拟微重力的旋转生物反应器从小鼠胚胎干细胞(MES)中生成功能性肝样细胞。在含有外源生长因子和激素的三维培养体系中，ES细胞可以分化为具有典型成熟肝细胞形态特征的肝样细胞。逆转录聚合酶链式反应检测、Western印迹检测、免疫染色和流式细胞仪分析表明，这些细胞在分化过程中表达肝脏特异的基因和蛋白。支架上的分化细胞进一步表现出肝细胞的形态特征和生物标志物，包括白蛋白产生、细胞色素P450活性和低密度脂蛋白摄取。当这些干细胞支架被移植到严重的联合免疫缺陷小鼠体内时，3D结构仍然有效，在体内经历了肝样细胞的进一步分化和成熟。总之，ES细胞在可生物降解的聚合物支架和旋转微重力生物反应器中的生长和分化可以产生对涉及生物人工肝和工程肝组织的研究有用的功能性和组织性肝细胞。
Embryonic stem (ES) cells are pluripotent cells that are capable of differentiating all the somatic cell lineages, including those in the liver tissue. We describe the generation of functional hepatic-like cells from mouse ES (mES) cells using a biodegradable polymer scaffold and a rotating bioreactor that allows simulated microgravity. Cells derived from ES cells cultured in the three-dimensional (3D) culture system with exogenous growth factors and hormones can differentiate into hepatic-like cells with morphologic characteristics of typical mature hepatocytes. Reverse-transcription polymerase chain-reaction testing, Western blot testing, immunostaining, and flow cytometric analysis show that these cells express hepatic-specific genes and proteins during differentiation. Differentiated cells on scaffolds further exhibit morphologic traits and biomarkers characteristic of liver cells, including albumin production, cytochrome P450 activity, and low-density lipoprotein uptake. When these stem cell-bearing scaffolds are transplanted into severe combined immunodeficient mice, the 3D constructs remained viable, undergoing further differentiation and maturation of hepatic-like cells in vivo. In conclusion, the growth and differentiation of ES cells in a biodegradable polymer scaffold and a rotating microgravity bioreactor can yield functional and organizational hepatocytes useful for research involving bioartificial liver and engineered liver tissue.