Evaluation the effect of the conditioned medium from human testicular sperm extraction cell cultures on in vitro maturation in mice

Background: It was shown previously, that conditioned medium from mouse testicular cells induces female germ cell development in embryonic stem cells (ESCs) in several mammals including mice and Buffalo. Lacham-Kaplan and co-workers claimed the formation of artificial ovarian structures containing oocyte-like structures following culture with testicular conditioned medium. Similar data using gene expression profile assessment was shown in Buffalo ESCs. These findings indicate that factors secreted by testicular cells may support in vitro maturation (IVM) in mammals.Objective: Here the impact of human testicular cell conditioned medium (TCCM) on mouse oocyte in vitro maturation was investigated.Materials and Methods: 240 germinal vesicle (GV) oocytes were obtained from NMRI mice, aged 4-6 weeks that injected 5IU PMSG 48 hours before. GV oocytes without cumulus cells were subjected to in vitro maturation in TCCM (test group; 120 GVs) and DMEM+10%FBS (control group; 120 GVs). GV maturation rate was checked by microscopic observation at 8, 16 and 24 hr following IVM in both groups. The MII (metaphase II) oocytes were subjected for IVF. The fertilization rate was evaluated after 24, 48 and 72 hr.Results: There was a significant difference between the maturation rates in TCCM (31.67%) and control (0%; p<0.05). IVF success rate for MII oocytes obtained in TCCM group was 28.94%.Conclusion: Our data confirms previous findings which shown there are supportive factors in TCCM for female germ cell development. For further investigations we aim to evaluate the effect of TCCM on human IVM and assess the metabolomics profile of TCCM in the future.

کلیدواژه ها:

Conditioned medium ، Germinal vesicle ، Testicular sperm extraction ، In vitro fertilization ، In vitro maturation