Leschik Julia, Stefanovic Sonia, Brinon Benjamin, Pucéat Michel
INSERM-Evry University UMR861, 5 Rue Desbruères, 91030 Evry, France.
Nat Protoc. 2008;3(9):1381-7. doi: 10.1038/nprot.2008.116.
Primate nonhuman and human embryonic stem (ES) cells provide a powerful model of early cardiogenesis. Furthermore, engineering of cardiac progenitors or cardiomyocytes from ES cells offers a tool for drug screening in toxicology or to search for molecules to improve and scale up the process of cardiac differentiation using high-throughput screening technology, as well as a source of cell therapy of heart failure. Spontaneous differentiation of ES cells into cardiomyocytes is, however, limited. Herein, we describe a simple protocol to commit both rhesus and human ES cells toward a cardiac lineage and to sort out early cardiac progenitors. Primate ES cells are challenged for 4 d with the cardiogenic morphogen bone morphogenetic protein 2 (BMP2) and sorted out using anti-SSEA-1 antibody-conjugated magnetic beads. Cardiac progenitor cells can be generated and isolated in 4 d using this protocol.
非人灵长类动物和人类胚胎干细胞为早期心脏发生提供了一个强大的模型。此外，从胚胎干细胞工程化生成心脏祖细胞或心肌细胞，为毒理学中的药物筛选提供了一种工具，或者利用高通量筛选技术寻找分子以改善和扩大心脏分化过程，同时也是心力衰竭细胞治疗的一个来源。然而，胚胎干细胞自发分化为心肌细胞是有限的。在此，我们描述了一种简单的方案，可使恒河猴和人类胚胎干细胞定向分化为心脏谱系，并筛选出早期心脏祖细胞。用心脏发生形态发生素骨形态发生蛋白2（BMP2）对灵长类胚胎干细胞进行4天的诱导，然后使用抗SSEA-1抗体偶联磁珠进行筛选。使用该方案可在4天内生成并分离出心脏祖细胞。
