人胚胎干细胞接种在鼠尾胶原包被培养板上定向分化为血管内皮细胞

背景：胚胎干细胞在体外可诱导分化为血管内皮细胞,进而形成血管,因而成为血管组织工程理想的种子细胞来源之一。 目的：探讨建立定向诱导人胚胎干细胞向血管内皮细胞分化的方法。 方法：在小鼠胚胎成纤维细胞饲养层上培养人胚胎干细胞H1,将H1细胞团块转移到鼠尾胶原包被的培养板,贴壁24- 48 h后,培养基换为含不同辅助因子和内皮细胞生长添加剂的EGM-2内皮细胞培养基。 结果与结论：①在EGM-2内皮细胞培养基诱导下,细胞逐步表达内皮细胞特异性标记基因VEGFR-2、PECAM1、vWF、CD34、VE-cadherin、GATA-2。②分化细胞表达内皮细胞特异性标记VE-cadherin、CD31。③分化细胞具有吞噬低密度脂蛋白功能。说明将人胚胎干细胞接种在鼠尾胶原包被培养板上,通过EGM-2内皮培养体系诱导,可直接分化为功能性血管内皮细胞。为研究胞外基质对诱导胚胎干细胞血管内皮细胞分化的作用以及相关信号分子机制打下了基础。

Human embryonic stem cells seeded in rat tail collagen-coated culture plates directly differentiate into vascular endothelial cells

BACKGROUND:Human embryonic stem cells (hESCs) can be induced to differentiate into vascular endothelial cells in vitro, which be involved in the formation of the blood vessels. Therefore, hESCs is one of the promised cell resources for vascular tissue engineering. OBJECTIVE:To establish a method to directly induce the hESCs to differentiate into vascular endothelial cells. METHODS:hESCs H1 was cultured on mouse embryonic fibroblast (MEF) in serum-free ESC medium. After passaged, H1 cell clumps were transferred to rat tail collagen-coated culture plates, and then cultured on EGM-2 endothelial medium added with endothelial cell growth supplements and growth factors after adherent cultured for 24-48 hours. RESULTS AND CONCLUSION:Cells cultured on EGM-2 endothelial medium gradually expressed endothelial-specific genes VEGFR-2, PECAM1, vWF, CD34, VE-cadherin and GATA-2. The differentiated cells could express endothelial- specific genes VE-cadherin and CD31. The differentiated cells could uptake low density lipoprotein. hESCs on tail collagen-coated culture plates could directly differentiate into vascular endothelial cells in EGM-2 endothelial medium. The establishment of endothelial differentiation system will lay a foundation for further researches on the effects of extracellular matrix on inducing hESCs to differentiate vascular endothelial cells and its molecular mechanisms.