title: Generation of cardiac and endothelial cells from neonatal mouse testis-derived multipotent germline stem cells.
abstract: 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.
authors.affiliation: Department of Pediatrics, Graduate School of Medicine, Kyoto University, 54 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan.
authors.forename: Shiro
authors.initials: S
authors.lastname: Baba
authors.forename: Toshio
authors.initials: T
authors.lastname: Heike
authors.forename: Katsutsugu
authors.initials: K
authors.lastname: Umeda
authors.forename: Toru
authors.initials: T
authors.lastname: Iwasa
authors.forename: Shinji
authors.initials: S
authors.lastname: Kaichi
authors.forename: Yoshimi
authors.initials: Y
authors.lastname: Hiraumi
authors.forename: Hiraku
authors.initials: H
authors.lastname: Doi
authors.forename: Momoko
authors.initials: M
authors.lastname: Yoshimoto
authors.forename: Mito
authors.initials: M
authors.lastname: Kanatsu-Shinohara
authors.forename: Takashi
authors.initials: T
authors.lastname: Shinohara
authors.forename: Tatsutoshi
authors.initials: T
authors.lastname: Nakahata
mesh_terms: 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
chemical_list: D015415:Biomarkers; D007473:Ion Channels; D040301:Vascular Endothelial Growth Factor Receptor-2