Establishment and characterization of CD-1 neonatal female germline stem cell lines / 中国组织工程研究

ABSTRACT

BACKGROUND:In traditional view, the germ cels in mammal ovaries were considered to lose the capacity of self-renew after birth. However, recent studies have showed the existence of female germline stem cels (FGSCs) which take the responsibility for regenerating oocytes in the ovaries of postnatal mammals. OBJECTIVE:To establish the FGSC line and to explore the biological characterization of the FGSC line from neonatal CD-1 mouse ovaries. METHODS:Using two-step enzymatic digestion and immunomagnetic purification, Fragilis-positive cels were isolated from neonatal CD-1 mice ovaries and subjected to long-term culture. Then their biological characteristics were studies with RT-PCR, fluorescent immunocytochemistry and cytogenetic analysis. RESULTS AND CONCLUSION:FGSCs from neonatal CD-1 mice aged 3-5 days were isolated and long-term cultured for 70 passages to establish a cellline. This cellline expressed Mvh, Dazl, Oct-4, Stela, Blimp1, Fragilis, and was proven to be mitoticaly active germ cels by BrdU/Fragilis dual immunofluorescence analysis. Moreover, cytogenetic analysis by Gimsa staining confirmed their normal karyotype. In al, these results indicate the existence of FGSCs with mitotic activity in neonatal CD-1 mouse ovaries.