In traditional view, the germ cells 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 cells (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 cells 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 cell line. This cell line expressed Mvh, Dazl, Oct-4, Stella, Blimp1, Fragilis, and was proven to be mitotically active germ cells by BrdU/Fragilis dual immunofluorescence analysis. Moreover, cytogenetic analysis by Gimsa staining confirmed their normal karyotype. In all, these results indicate the existence of FGSCs with mitotic activity in neonatal CD-1 mouse ovaries.RESULTS AND CONCLUSION:
Funding: the National Basic Research Program of China, No. 2013CB967401; the National Natural Science Foundation of China, No. 81370675RESULTS AND CONCLUSION:
Bi HW, Wu J. Establishment and characterization of CD-1 neonatal female germline stem cell lines. Zhongguo Zuzhi Gongcheng Yanjiu. 2014;18(19):3088-3093.