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Tissue-Specific Stem Cells |
1 Department of Pediatrics, Graduate School of Medicine, Kyoto University, Kyoto, Japan
2 Department of Molecular Genetics, Graduate School of Medicine, Kyoto University, Kyoto, Japan
* To whom correspondence should be addressed. E-mail: tnakaha{at}kuhp.kyoto-u.ac.jp.
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Abstract |
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Multipotent germline stem (mGS) cells have been established from neonatal mouse testes. Here we compared mGS, embryonic stem (ES) cells 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, about 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 the Flk1+ ES cells, while the Flk1+ EG cells generated equivalent numbers as the Flk1+ mGS cells. 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 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.
Key Words. Mouse, Germline, Somatic Stem Cells, Pluripotent Stem Cells, Embryonic Stem Cell, In vitro Differentiation
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