Lagarkova Maria A, Volchkov Pavel Y, Philonenko Elena S, Kiselev Sergei L
Vavilov Institute of General Genetics, Russian Academy of Science, Moscow, Russia.
Cell Cycle. 2008 Sep 15;7(18):2929-35. doi: 10.4161/cc.7.18.6700.
Human embryonic stem cells (hESCs) are to be considered as a valuable source for regenerative medicine because of their capacity to differentiate into all cell types. We have developed an efficient culture system to differentiate hECSs into endothelial cells without the formation of embryoid bodies Establishing appropriate culture conditions with a cocktail of growth factors allowed us to differentiate hESCs directly to endothelial primary culture with about 50% efficiency. CD31 immunomagnetic cell sorting was used to purify derived endothelium from the primary culture of hESCs. Isolated endothelial cells expressed immunological markers (vWF, CD105), specific genes (VE-cadherin, KDR, GATA-2, GATA-3, eNOS), and formed cord-like structures on collagen matrix and in Matrigel assay. During differentiation to endothelial lineage promoter regions of the genes involved in specific cell fate determination and homeostasis (GATA-2,-3, and eNOS) underwent intensive hypomethylation which correlated with the gene expression. Overall our data demonstrate that direct differentiation of hESCs leads to endothelial cells that acquire epigenetic patterning similar to the functional endothelial cells of the organism.
人类胚胎干细胞（hESCs）因其能够分化为所有细胞类型，而被视为再生医学的宝贵来源。我们开发了一种高效的培养系统，可将hESCs分化为内皮细胞，且不会形成胚状体。通过使用生长因子混合物建立合适的培养条件，我们能够以约50%的效率将hESCs直接分化为内皮原代培养物。利用CD31免疫磁珠细胞分选技术从hESCs的原代培养物中纯化衍生的内皮细胞。分离出的内皮细胞表达免疫标记物（vWF、CD105）、特定基因（VE-钙黏蛋白、KDR、GATA-2、GATA-3、eNOS），并在胶原基质和基质胶试验中形成条索状结构。在向内皮谱系分化过程中，参与特定细胞命运决定和内环境稳定的基因（GATA-2、-3和eNOS）的启动子区域发生了强烈的低甲基化，这与基因表达相关。总体而言，我们的数据表明，hESCs的直接分化产生的内皮细胞获得了与机体功能性内皮细胞相似的表观遗传模式。
