Isolation and enrichment of mouse female germ line stem cells

Khosravi-Farsani, S.; Fardin, A.; Habibi Roudkenar, M.; Sobhani, A.
June 2014
Iranian Journal of Reproductive Medicine;Jun2014 Supplement, Vol. 12, p30
Academic Journal
Introduction: 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 female germ line stem cell enrichment and establishing the best procedure. Materials and Methods: After digesting neonate ovary from C57B1/6 mice, we performed 2 different isolation experiments: Magnetic activating cell sorting (MACS) and preplating. MACS was done by two different antibodies against MVH and SSEA1 markers. Then to characterize colony forming cells by RT-PCR (for analysis of expression of Oct4, Nanog, C-kit, Fragilis, Mvh, Dazl, Scp3 andZp3), AP activity test and Immunocytochemistry, they were passaged and proliferated in vitro. Results: Data showed that colonies can be seen more frequently in preplating technique than that in MACs. Using the SSEA1 antibody with MACS, 1.98±0.49% (Mean±SD) positive cells were yield as compared to the total cells sorted. The colonies formed after preplating expressed pluripotency and germ stem cell markers (Oct4, Nanog, C-kit, Fragilis, Mvh and Dazl) but did not express Zp3 and Sep3 at the mRNA level. Immunocytochemistry in these colonies further confirmed the presence of OCT4 and MVH proteins and alkaline phosphatase activity measured by AP-kit showed positive reaction. Conclusion: We established a simple and efficient preplating technique that allows culture and enrichment of female germ line stem cells from neonatal mouse ovaries.


Related Articles

  • Intense Sperm-Mediated Sexual Conflict Promotes Reproductive Isolation in Caenorhabditis Nematodes. Ting, Janice J.; Woodruff, Gavin C.; Leung, Gemma; Shin, Na-Ra; Cutter, Asher D.; Haag, Eric S. // PLoS Biology;Jul2014, Vol. 12 Issue 7, p1 

    Sperm from other species invade female tissues to cause sterility and death, helping to keep nematode species boundaries intact.

  • Genetic Incompatibility Dampens Hybrid Fertility More Than Hybrid Viability: Yeast as a Case Study. Meibo Xu; Xionglei He // PLoS ONE;2011, Vol. 6 Issue 4, p1 

    Genetic incompatibility is believed to be the major cause of postzygotic reproductive isolation. Despite huge efforts seeking for speciation-related incompatibilities in the past several decades, a general understanding of how genetic incompatibility evolves in affecting hybrid fitness is not...

  • Isolation and molecular identification of mycoplasma genitalium from the secretion of genital tract in infertile male and female. Moghadam, Naeime Mohseni; Kheirkhah, Babak; Mirshekari, Toraj Reza; Harandi, Majid Fasihi; Tafsiri, Elham // Iranian Journal of Reproductive Medicine;Sep2014, Vol. 12 Issue 9, p601 

    Background: Mycoplasmas can cause acute and chronic diseases at multiple sites with wide-range complications and have been implicated as cofactors in diseases. The infections influenced form genital mycoplasmas specifically Mycoplasma hominis and Mycoplasma genitalium potentially affect...

  • The uncharacterized gene 1700093K21Rik and flanking regions are correlated with reproductive isolation in the house mouse, Mus musculus. Kass, David; Janoušek, Václav; Wang, Liuyang; Tucker, Priscilla // Mammalian Genome;Jun2014, Vol. 25 Issue 5/6, p223 

    Reproductive barriers exist between the house mouse subspecies, Mus musculus musculus and M. m. domesticus, members of the Mus musculus species complex, primarily as a result of hybrid male infertility, and a hybrid zone exists where their ranges intersect in Europe. Using single nucleotide...

  • The Pace of Hybrid Incompatibility Evolution in House Mice. Wang, Richard J.; White, Michael A.; Payseur, Bret A. // Genetics;9/1/2015, Vol. 201 Issue 1, p229 

    Hybrids between species are often sterile or inviable. This form of reproductive isolation is thought to evolve via the accumulation of mutations that interact to reduce fitness when combined in hybrids. Mathematical formulations of this "Dobzhansky-Muller model" predict an accelerating buildup...

  • Patterns of Reproductive Isolation in Toads. Malone, John H.; Fontenot, Brian E. // PLoS ONE;2008, Vol. 3 Issue 12, p1 

    Understanding the general features of speciation is an important goal in evolutionary biology, and despite significant progress, several unresolved questions remain. We analyzed an extensive comparative dataset consisting of more than 1900 crosses between 92 species of toads to infer patterns of...

  • Multifaceted, Cross-Generational Costs of Hybridization in Sibling Drosophila species. Myers, Erin M.; Harwell, Tiffany I.; Yale, Elizabeth L.; Lamb, Abigail M.; Frankino, W. Anthony // PLoS ONE;Nov2013, Vol. 8 Issue 11, p1 

    Maladaptive hybridization, as determined by the pattern and intensity of selection against hybrid individuals, is an important factor contributing to the evolution of prezygotic reproductive isolation. To identify the consequences of hybridization between Drosophila pseudoobscura and D....

  • Characterization of Prdm9 in Equids and Sterility in Mules. Steiner, Cynthia C.; Ryder, Oliver A. // PLoS ONE;Apr2013, Vol. 8 Issue 4, p1 

    : Prdm9 (Meisetz) is the first speciation gene discovered in vertebrates conferring reproductive isolation. This locus encodes a meiosis-specific histone H3 methyltransferase that specifies meiotic recombination hotspots during gametogenesis. Allelic differences in Prdm9, characterized for a...

  • Erratum to: Isolation of Sporothrix schenckii From the Claws of Domestic Cats (Indoor and Outdoor) and in Captivity in São Paulo (Brazil). Borges, Tatiana; Rossi, Claudio; Fedullo, José; Taborda, Carlos; Larsson, Carlos // Mycopathologia;Oct2013, Vol. 176 Issue 3/4, p317 

    A correction to the article "Isolation of Sporothrix schenckii From the Claws of Domestic Cats (Indoor and Outdoor) and in Captivity in São Paulo (Brazil)" that was published in the August 15, 2013 issue is presented.


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics