Generation of cardiac and endothelial cells from neonatal mouse testis-derived multipotent germline stem cells

被引:36
作者
Baba, Sbiro
Heike, Toshio
Umeda, Katsutsugu
Iwasa, Toru
Kaichi, Shinji
Hiraumi, Yoshiml
Doi, Hiraku
Yoshimoto, Momoko
Kanatsu-Shinohara, Mito
Shinohara, Takashi
Nakahata, Tatsutosbi
机构
[1] Kyoto Univ, Grad Sch Med, Dept Pediat, Sakyo Ku, Kyoto 6068507, Japan
[2] Kyoto Univ, Grad Sch Med, Dept Mol Genet, Sakyo Ku, Kyoto 6068507, Japan
关键词
mouse; germline; somatic stem cells; pluripotent stem cells; embryonic stem cell; in vitro differentiation;
D O I
10.1634/stemcells.2006-0574
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Multipotent germline stem (mGS) cells have been established from neonatal mouse testes. Here, we compared mGS, embryonic stem (ES), and embryonic germ (EG) cells with regard to their ability to differentiate into mesodermal cells, namely, cardiomyocytes and endothelial cells. The in situ morphological appearances of undifferentiated mGS, ES, and EG cells were similar, and 4 days after being induced to differentiate, approximately 30%-40% of each cell type differentiated into Flk1(+) cells. The sorted Flk1(+) cells differentiated efficiently into cardiomyocytes and endothelial cells. By day 10 after differentiation induction, the three cell types generated equal number of endothelial colonies. However, by day 13 after differentiation induction, the Flk1(+) mGS cells generated more contractile colonies than did the Flk1(+) ES cells, whereas the Flk1(+) EG cells generated equivalent numbers as the Flk1(+) mGS cells. Reverse transcriptase polymerase chain reaction (RT-PCR) analysis of differentiation markers such as Rex1, FGF-5, GATA-4, Brachyury, and Flk1 revealed that mGS cells expressed these markers more slowly during days 0-4 after differentiation induction than did ES cells, but that this mGS cell pattern was similar to that of the EG cells. RT-PCR analysis also revealed that the three differentiation cell types expressed various cardiac markers. Moreover, immunohistochemical analysis revealed that the contractile colonies derived from Flk1(+) mGS cells express mature cardiac cell-specific markers. In conclusion, mGS cells are phenotypically similar to ES and EG cells and have a similar potential to differentiate into cardiomyocytes and endothelial cells.
引用
收藏
页码:1375 / 1383
页数:9
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