Generation of cardiac and endothelial cells from neonatal mouse testis-derived multipotent germline stem cells.
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. Disclosure of potential conflicts of interest is found at the end of this article.
Baba
Shiro
S
Department of Pediatrics, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
Heike
Toshio
T
Umeda
Katsutsugu
K
Iwasa
Toru
T
Kaichi
Shinji
S
Hiraumi
Yoshimi
Y
Doi
Hiraku
H
Yoshimoto
Momoko
M
Kanatsu-Shinohara
Mito
M
Shinohara
Takashi
T
Nakahata
Tatsutoshi
T
D000818:Animals; D000831:Animals, Newborn; D015415:Biomarkers / Q000032:analysis; D002454:Cell Differentiation; D002478:Cells, Cultured; D042783:Endothelial Cells / Q000166:cytology*; D005854:Germ Cells / Q000166:cytology*; D007473:Ion Channels / Q000032:analysis; D008297:Male; D008564:Membrane Potentials / Q000502:physiology; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology*; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000166:cytology*; D013737:Testis / Q000166:cytology*; D040301:Vascular Endothelial Growth Factor Receptor-2 / Q000378:metabolism
D015415:Biomarkers; D007473:Ion Channels; D040301:Vascular Endothelial Growth Factor Receptor-2