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

Shiro Baba, Toshio Heike, Katsutsugu Umeda, Toru Iwasa, Shinji Kaichi, Yoshimi Hiraumi, Hiraku Doi, Momoko Yoshimoto, Mito Kanatsu-Shinohara, Takashi Shinohara, Tatsutoshi Nakahata

Research output: Contribution to journalArticle

37 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)1375-1383
Number of pages9
JournalSTEM CELLS
Volume25
Issue number6
DOIs
StatePublished - Jun 1 2007

Keywords

  • Embryonic stem cell
  • Germline
  • In vitro differentiation
  • Mouse
  • Pluripotent stem cells
  • Somatic stem cells

ASJC Scopus subject areas

  • Molecular Medicine
  • Developmental Biology
  • Cell Biology

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