Liu Yu-xiao, Yue Wen, Ji Lei, Nan Xue, Pei Xue-tao
Stem Cell and Regenerative Medicine Lab, Beijing Institution of Transfusion Medicine, Beijing 100850, China.
BMC Dev Biol. 2010 Aug 10;10:85. doi: 10.1186/1471-213X-10-85.
We recently developed a new method to induce human stem cells (hESCs) differentiation into hematopoietic progenitors by cell extract treatment. Here, we report an efficient strategy to generate erythroid progenitors from hESCs using cell extract from human fetal liver tissue (hFLT) with cytokines. Human embryoid bodies (hEBs) obtained of human H1 hESCs were treated with cell extract from hFLT and co-cultured with human fetal liver stromal cells (hFLSCs) feeder to induce hematopoietic cells. After the 11 days of treatment, hEBs were isolated and transplanted into liquid medium with hematopoietic cytokines for erythroid differentiation. Characteristics of the erythroid cells were analyzed by flow cytometry, Wright-Giemsa staining, real-time RT-PCR and related functional assays.
The erythroid cells produced from hEBs could differentiate into enucleated cells and expressed globins in a time-dependent manner. They expressed not only embryonic globins but also the adult-globin with the maturation of the erythroid cells. In addition, our data showed that the hEBs-derived erythroid cells were able to act as oxygen carriers, indicating that hESCs could generate functional mature erythroid cells.
Cell extract exposure with the addition of cytokines resulted in robust erythroid -like differentiation of hEBs and these hEBs-derived erythroid cells possessed functions similar to mature red blood cells.
我们最近开发了一种通过细胞提取物处理诱导人类干细胞(hESCs)分化为造血祖细胞的新方法。在此,我们报告一种利用人胎肝组织(hFLT)细胞提取物与细胞因子从hESCs生成红系祖细胞的有效策略。将人H1 hESCs获得的人胚状体(hEBs)用hFLT细胞提取物处理,并与人胎肝基质细胞(hFLSCs)饲养层共培养以诱导造血细胞。处理11天后,分离hEBs并移植到含有造血细胞因子的液体培养基中进行红系分化。通过流式细胞术、瑞氏-吉姆萨染色、实时RT-PCR及相关功能测定分析红系细胞的特征。
hEBs产生的红系细胞可分化为无核细胞,并以时间依赖性方式表达珠蛋白。随着红系细胞的成熟,它们不仅表达胚胎珠蛋白,还表达成人珠蛋白。此外,我们的数据表明,hEBs来源的红系细胞能够作为氧载体,这表明hESCs可以产生功能性成熟红系细胞。
添加细胞因子的细胞提取物暴露导致hEBs强烈的类红系分化,且这些hEBs来源的红系细胞具有与成熟红细胞相似的功能。