title: Differentiation of spermatogonial stem cell-like cells from murine testicular tissue into haploid male germ cells in vitro.
abstract: In vitro differentiation of spermatogonial stem cells (SSCs) promotes the understanding of the mechanism of spermatogenesis. The purpose of this study was to isolate spermatogonial stem cell-like cells from murine testicular tissue, which then were induced into haploid germ cells by retinoic acid (RA). The spermatogonial stem cell-like cells were purified and enriched by a two-step plating method based on different adherence velocities of SSCs and somatic cells. Cell colonies were present after culture in M1-medium for 3 days. Through alkaline phosphatase, RT-PCR and indirect immunofluorescence cell analysis, cell colonies were shown to be SSCs. Subsequently, cell colonies of SSCs were cultured in M2-medium containing RA for 2 days. Then the cell colonies of SSCs were again cultured in M1-medium for 6-8 days, RT-PCR and indirect immunofluorescence cell analysis were chosen to detect haploid male germ cells. It could be demonstrated that 10(-7) mol l(-1) of RA effectively induced the SSCs into haploid male germ cells in vitro.
authors.affiliation: College of Animal Science and Technology, Northwest A&F University, Yangling, 712100, Shaanxi, People's Republic of China.
authors.forename: Peng
authors.initials: P
authors.lastname: Wang
authors.forename: Li-Juan
authors.initials: LJ
authors.lastname: Suo
authors.forename: Hua
authors.initials: H
authors.lastname: Shang
authors.forename: Ying
authors.initials: Y
authors.lastname: Li
authors.forename: Guang-Xuan
authors.initials: GX
authors.lastname: Li
authors.forename: Qing-Wang
authors.initials: QW
authors.lastname: Li
authors.forename: Jian-Hong
authors.initials: JH
authors.lastname: Hu