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Cryopreserved ovine spermatogonial stem cells maintain stemness and colony forming ability in vitro

Asian pacific Journal of Reproduction 2023年第6期12卷 273-280页

作者： R.Kumar Pramod Deepthi Varughese A.Javed Jameel bhisma narayan panda Soma Goswami Abhijit Mitra ICMR-National Animal Resource Facility for Biomedical Research HyderabadTelangana500101India Inter University Centre for Biomedical Research&Super Speciality Hospital Kerala686009India College of Veterinary and Animal Sciences MannuthyKerala680651India Translational Health Science and Technology Institute FaridabadHaryana121001India G V Research Platform HyderabadTelangana500101India Department of Animal Husbandry&Dairying MoFAHDKrishi BhavanNew Delhi110001India

Objective:To assess the effect of cryopreservation on stemness and proliferation potential of sheep spermatogonial stem cells(SSCs)in ***:Sheep testicular cells were isolated and putative SSCs were enriched by the laminin-based differential plating *** SSCs were co-cultured with the Sertoli cell feeder prepared by the Datura Stramonium Agglutinin(DSA-lectin)-based *** cultured putative SSCs were cryopreserved in Dulbecco's Modified Eagle Medium-10%fetal bovine serum mixture(DMEM-10%FBS)media containing 10%dimethyl sulfoxide(DMSO)alone or 10%DMSO plus 200 mM *** putative SSCs were evaluated for their proliferation potential using in vitro culture and stemness by ***,the transfection ability of cryopreserved putative SSCs was ***:We isolated 91%viable testicular cells from sheep *** majority of the laminin enriched cells expressed the SSC related marker,***-culture of sheep putative SSCs with Sertoli cell feeder resulted in the generation of stable colonies,and the expression of SSC marker was maintained after several passages.A significantly higher number of viable putative SSCs was recovered from SSCs cryopreserved in media containing 10%DMSO and 200 mM trehalose compared to 10%DMSO alone(P<0.01).Cryopreserved putative SSCs formed colonies and showed SSC marker expression similar to the non-cryopreserved putative *** appearance of green fluorescent colonies over the Sertoli cell feeder indicated that cryopreserved sheep SSCs were successfully ***:Cryopreserved putative SSCs can retain their stemness,colony forming ability,and transfection efficiency in *** research may help in the effective preservation of germplasm and the generation of transgenic ovine species.

关键词： Spermatogonial stem cell Sheep Stemness Cryopreservation Transfection

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