Kurita Ryo, Sasaki Erika, Yokoo Tomoko, Hiroyama Takashi, Takasugi Kashiya, Imoto Hideyuki, Izawa Kiyoko, Dong Yan, Hashiguchi Takao, Soda Yasushi, Maeda Toyoki, Suehiro Youko, Tanioka Yoshikuni, Nakazaki Yukoh, Tani Kenzaburo
Department of Molecular Genetics, Division of Molecular and Clinical Genetics, Medical Institute of Bioregulation, Kyushu University, 3-1-1, Maidashi, Fukuoka 812-8582, Japan.
Stem Cells. 2006 Sep;24(9):2014-22. doi: 10.1634/stemcells.2005-0499. Epub 2006 May 25.
The development of embryonic stem cell (ESC) therapies requires the establishment of efficient methods to differentiate ESCs into specific cell lineages. Here, we report the in vitro differentiation of common marmoset (CM) (Callithrix jacchus) ESCs into hematopoietic cells after exogenous gene transfer using vesicular stomatitis virus-glycoprotein-pseudotyped lentiviral vectors. We transduced hematopoietic genes, including tal1/scl, gata1, gata2, hoxB4, and lhx2, into CM ESCs. By immunochemical and morphological analyses, we demonstrated that overexpression of tal1/scl, but not the remaining genes, dramatically increased hematopoiesis of CM ESCs, resulting in multiple blood-cell lineages. Furthermore, flow cytometric analysis demonstrated that CD34, a hematopoietic stem/progenitor cell marker, was highly expressed in tal1/scl-overexpressing embryoid body cells. Similar results were obtained from three independent CM ESC lines. These results suggest that transduction of exogenous tal1/scl cDNA into ESCs is a promising method to induce the efficient differentiation of CM ESCs into hematopoietic stem/progenitor cells.
胚胎干细胞(ESC)疗法的发展需要建立有效的方法将胚胎干细胞分化为特定的细胞谱系。在此,我们报告了使用水泡性口炎病毒糖蛋白假型慢病毒载体进行外源基因转移后,普通狨猴(CM)(绢毛猴)胚胎干细胞在体外分化为造血细胞的情况。我们将包括tal1/scl、gata1、gata2、hoxB4和lhx2在内的造血基因转导到CM胚胎干细胞中。通过免疫化学和形态学分析,我们证明tal1/scl的过表达而非其余基因,显著增加了CM胚胎干细胞的造血功能,产生了多种血细胞谱系。此外,流式细胞术分析表明,造血干细胞/祖细胞标志物CD34在过表达tal1/scl的胚状体细胞中高度表达。从三个独立的CM胚胎干细胞系获得了类似结果。这些结果表明,将外源tal1/scl cDNA转导到胚胎干细胞中是一种诱导CM胚胎干细胞有效分化为造血干细胞/祖细胞的有前景的方法。