Sequential analysis of α- and β-globin gene expression during erythropoietic differentiation from primate embryonic stem cells

被引:12
作者
Umeda, Katsutsugu
Heike, Toshio
Nakata-Hizume, Mami
Niwa, Akira
Arai, Masato
Shinoda, Gen
Ma, Feng
Suemori, Hirofumi
Luo, Hong Yuan
Chui, David H. K.
Torii, Ryuzo
Shibuya, Masabumi
Nakatsuji, Norio
Nakahata, Tatsutoshi
机构
[1] Kyoto Univ, Dept Pediat, Grad Sch Med, Sakyo Ku, Kyoto 6068507, Japan
[2] Kyoto Univ, Lab Embryon Stem Cell Res, Stem Cell Res Ctr, Inst Frontier Med Sci, Kyoto 6068507, Japan
[3] Boston Univ, Sch Med, Dept Med, Boston, MA 02118 USA
[4] Shiga Univ Med Sci, Res Ctr Anim Life Sci, Otsu, Shiga, Japan
[5] Univ Tokyo, Inst Med Sci, Div Genet, Tokyo, Japan
[6] Kyoto Univ, Dept Dev & Differentiat, Inst Frontier Sci, Kyoto 6068507, Japan
关键词
embryonic stem cells; erythroid progenitors;
D O I
10.1634/stemcells.2006-0199
中图分类号
Q813 [细胞工程];
学科分类号
摘要
The temporal pattern of embryonic, fetal, and adult globin expression in the alpha (zeta ->alpha) and beta (epsilon ->gamma and gamma ->beta) clusters were quantitatively analyzed at the transcriptional and translational levels in erythrocytes induced from primate embryonic stem cells in vitro. When vascular endothelial growth factor receptor-2(high) CD34(+) cells were harvested and reseeded onto OP9 stromal cells, two-wave erythropoiesis occurred sequentially. Immunostaining and real-time reverse transcription-polymerase chain reaction analyses of floating mature erythrocytes revealed that globin switches occurred in parallel with the erythropoietic transition. Colony-forming assays showed replacement of primitive clonogenic progenitor cells with definitive cells during culturing. A decline in embryonic zeta- and epsilon-globin expression at the translational level occurred in individual definitive erythroid progenitors. Expression of beta-globin in individual definitive erythroid progenitors was upregulated in the presence of OP9 stromal cells. Thus, this system reproduces early hematopoietic development in vitro and can serve as a model for analyzing the mechanisms of the globin switch in humans.
引用
收藏
页码:2627 / 2636
页数:10
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