Direct Differentiation of Homogeneous Human Adipose Stem Cells Into Functional Hepatocytes by Mimicking Liver Embryogenesis
Direct Differentiation of Homogeneous Human Adipose Stem Cells Into Functional Hepatocytes by Mimicking Liver Embryogenesis
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通过模仿肝胚胎发生将同质人脂肪干细胞直接分化为功能性肝细胞
DOI：
10.1002/jcp.24501
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发表时间：
2014-06-01
影响因子：
5.6
通讯作者：
Zhang, Haiyan
中科院分区：
文献类型：
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作者：
Li, Xueyang;Yuan, Jie;Zhang, Haiyan
成人脂肪组织干细胞（hASC）分化为肝细胞的潜力已经产生了许多兴奋的hASC在治疗应用中的可能用途。对hASCs向肝细胞可塑性的分子机制的理解将有助于使这种可能性成为现实。在本文中，我们显示了以高水平的CD 73、CD 90和CD 105为特征的hASC的同质群体在增殖条件下表达多能转录因子OCT 4、SOX 2、NANOG和SALL 4。高水平的激活素A允许hASC获得定形内胚层（DE）细胞的命运并同步表达特异性转录因子HEX、FOXA 2、SOX 17和GATA 4。使用通过模拟肝胚胎发生的可再现的三阶段方法，将hASC定向分化为功能性肝细胞。在第一阶段，通过在无血清培养基中培养2天和激活素A处理3天，将hASC诱导为DE细胞。接下来，在培养基中存在成纤维细胞生长因子（FGF）4和骨形态发生蛋白（BMP）2 5天诱导DE细胞的有效肝分化。通过依次暴露于肝细胞生长因子（HGF）、制瘤素M（OSM）和地塞米松（DEX）进一步成熟10天后，hASC衍生的肝细胞表达成熟肝细胞标志物并表现出功能特征，包括白蛋白分泌、糖原储存、尿素产生、药物转运蛋白活性和细胞色素P450活性。这些发现将有助于hASC衍生的肝细胞在治疗目的，代谢分析，药物毒性筛选和肝细胞功能研究中的应用。J.细胞。229：801-812，2014。© 2013 Wiley Periodicals，Inc.
The potential of adult human adipose tissue stem cells (hASCs) to differentiate into hepatocytes has generated much excitement over the possible use of hASCs in therapeutic applications. An understanding of the molecular mechanisms that underlie the plasticity of hASCs toward hepatocytes will help to make this possibility a reality. Herein, we show that a homogenous population of hASCs characterized by a high level of CD73, CD90, and CD105 express the pluripotent transcription factors OCT4, SOX2, NANOG, and SALL4 under proliferation conditions. A high level of activin A allows for hASCs acquiring the fate of definitive endoderm (DE) cells and expressing the specific transcription factors HEX, FOXA2, SOX17, and GATA4 synchronously. Using a reproducible three‐stage method by mimicking liver embryogenesis, hASCs were directed to differentiate into functional hepatocytes. In the first stage, hASCs were induced to become DE cells by 2 days cultured in serum‐free medium and 3 days of activin A treatment. Next, the presence of fibroblast growth factor (FGF) 4 and bone morphogenetic protein (BMP) 2 in the medium for 5 days induced efficient hepatic differentiation from DE cells. After 10 days of further maturated by the sequential exposure to hepatocyte growth factor (HGF), oncostatin M (OSM), and dexamethasone (DEX), the hASC‐derived hepatocytes expressed mature hepatocytes marker and exhibited functional characterization, including albumin secretion, glycogen storage, urea production, activity of drug transporters, and cytochrome P450 activity. These findings will be useful for the implementation of hASC‐derived hepatocytes in therapeutic purposes, metabolic analyses, drug toxicity screening, and studies of hepatocyte function. J. Cell. Physiol. 229: 801–812, 2014. © 2013 Wiley Periodicals, Inc.