Department of Infectious Diseases, Southwest Hospital, Third Military Medical University , Chongqing, People's Republic of China .
Tissue Eng Part A. 2010 Apr;16(4):1115-22. doi: 10.1089/ten.TEA.2009.0391.
Embryonic stem cells (ESCs) can have unlimited proliferation potential and the capability to differentiate into almost all types of tissues. The use of ESCs as a novel source of hepatocytes has therefore recently received much attention. To date, however, the efficient and mass differentiation of ESCs into hepatocytes in vitro has been proven to be difficult. In this study, we investigated a three-dimensional (3D) mouse ESC hepatic differentiation system. Cells derived from 5-day-old embryoid bodies were mixed with Matrigel and immediately seeded in a biodegradable polymer scaffold composed of poly-l-lactic acid and polyglycolic acid. Exogenous growth factors and hormones were added sequentially for 20 days to promote hepatic histogenesis. The system enabled spontaneous spheroid formation and mass cultivation of embryoid body-derived cells. The 3D differentiated hepatocyte-like cells were able to express several liver-specific markers and proteins, secrete albumin, store glycogen, and allow the uptake of low-density lipoproteins. These findings suggest that this supportive 3D environment with appropriate inducing factors could provide mouse ESC-derived cells with an environment closely mimicking that of hepatogenesis in vivo.
胚胎干细胞（ESCs）具有无限的增殖潜力和分化为几乎所有类型组织的能力。因此，最近人们对将 ESCs 用作新型肝细胞来源给予了极大关注。然而，迄今为止，已经证明在体外高效、大规模地将 ESCs 分化为肝细胞是困难的。在这项研究中，我们研究了一种三维（3D）小鼠 ESC 肝分化系统。将源自 5 天龄胚体的细胞与 Matrigel 混合，并立即接种到由聚 L-乳酸和聚乙二醇酸组成的可生物降解聚合物支架上。在接下来的 20 天中，添加外源性生长因子和激素以促进肝组织发生。该系统能够自发形成球体并大规模培养胚体衍生细胞。3D 分化的类肝细胞样细胞能够表达几种肝脏特异性标志物和蛋白质，分泌白蛋白，储存糖原，并允许摄取低密度脂蛋白。这些发现表明，这种具有适当诱导因子的支持性 3D 环境可以为小鼠 ESC 衍生细胞提供一种与体内肝发生非常相似的环境。
