Occupational Disease Research Center, Peking University Third Hospital, People's Republic of China.
Exp Cell Res. 2013 Jan 1;319(1):21-34. doi: 10.1016/j.yexcr.2012.09.009. Epub 2012 Oct 3.
Human embryonic stem (hES) cells can differentiate into cells of the three germ layers in vitro and serve as a powerful resource to study mechanisms involved in cell fate decisions. However, it is difficult to promote the directed and efficient differentiation of hES cells toward a specific lineage. Here we establish a stepwise strategy for generating hemato-endothelial and cardiac precursors from hES cells in single culture conditions. The efficiency of committing hES cells to three cell lineages was significantly higher with our approach than with exposure to single or multiple cytokines. Efficiency was determined using quantitative analysis by gene expression, flow cytometry, and colony assays. Several cytokines were sufficient to drive the efficient differentiation of hES cells into specific lineages. Each of these factors appeared to regulate specific steps of differentiation: BMP4 promoted the efficient formation of mesoderm; bFGF induced the differentiation of these mesodermal precursors to the hemangioblast fate; VEGF and TPO were required for the production of committed hematopoietic progenitors. This stepwise control of differentiation in vitro leads to a high frequency of hemato-endothelial and cardiac precursors derived from hES cells and offers a unique model for studying the molecular and cellular events that regulate hematopoiesis and cardiogenesis.
人类胚胎干细胞（hES）可在体外分化为三个胚层的细胞，是研究细胞命运决定机制的有力资源。然而，很难促进 hES 细胞向特定谱系的定向和有效分化。本文建立了一种在单一培养条件下从 hES 细胞生成造血内皮和心脏前体细胞的分步策略。与单一或多种细胞因子暴露相比，我们的方法显著提高了 hES 细胞向三个谱系定向分化的效率。通过基因表达、流式细胞术和集落分析的定量分析确定效率。几种细胞因子足以驱动 hES 细胞向特定谱系的有效分化。这些因子中的每一个似乎都调节分化的特定步骤：BMP4 促进中胚层的有效形成；bFGF 诱导这些中胚层前体向成血管球命运分化；VEGF 和 TPO 是产生定向造血祖细胞所必需的。这种体外分化的分步控制导致了高频率的造血内皮和心脏前体细胞来源于 hES 细胞，并为研究调节造血和心脏发生的分子和细胞事件提供了独特的模型。
