I see a high expression of X protein in spermatogonia and primary spermatocytes and want to check its role in spermatogenesis. For this I propose to isolate spermatoginia from normal mouse testis, knock my protein down via shRNA and re-inject it back into a sterile mice.
I want to know if isolating, growing spermatogonia and genetically manipulating it is possible as mentioned above. I will be very grateful if you can lend me some advice or protocols.
No its is not that easy to culture the spermatogonial cells in vitor and mantain them without differentiation... the alternate is do a in vivo testicular injection of your shRNA, identify the prefoundewrs and founders that carry your transgene (shRNA) .. directly isolate the spermatogonial cells and transplant...
Here are two papers that describe the testicular transgenesis protocols http://www.ncbi.nlm.nih.gov/pubmed/18552853
http://www.ncbi.nlm.nih.gov/pubmed/21760937
We are a Nanotechnology company that has developed fertility treatments for sperm. We collect and deplete moribund sperm and enrich for viable sperm and or Gender. We have worked with a variety of species; bovine,Equine,porcine,human, canine dolphins, etc. . Let me know your interest in working with us.We are looking for partners to help optimize this technology as well as other aspects of fertility. [Fetal cell isolation and enrichment]
Henry
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