Isolation and Enrichment of Mouse Female Germ
Line Stem Cells
The existence of female germ-line stem cells (FGSCs) has been the subject of a wide range of recent studies. Successful isolation and culture of FGSCs could facilitate studies on regenerative medicine and infertility treatments in the near future. Our aim in the present study was evaluation of the most commonly used techniques in enrichment of FGSCs and in establishment of the best procedure.
Materials and Methods
In this experimental study, after digesting neonate ovary from
C57Bl/6 mice, we performed 2 different isolation experiments: magnetic activated cell
sorting (MACS) and pre-plating. MACS was applied using two different antibodies against
mouse vasa homolog (MVH) and stage-specific embryonic antigen-1 (SSEA1) markers.
After the cells were passaged and proliferated
Data showed that colonies can be seen more frequently in pre-plating technique
than that in MACS. Using the SSEA1 antibody with MACS, 1.98 ± 0.49% (Mean ± SDV)
positive cells were yield as compared to the total cells sorted. The colonies formed after
pre-plating expressed pluripotency and germ stem cell markers (
We established a simple and an efficient pre-plating technique to culture and to enrich FGSCs from neonatal mouse ovaries.