Animals ; Cdc20 Proteins/metabolism* ; Cell Line ; Embryo, Mammalian/embryology* ; Embryonic Development ; Female ; Infertility, Female/metabolism* ; Mice ; Mutation ; Oocytes/metabolism*

Objective:To evaluate the cryopreservation effect of the home-made Strawtop micro-carrier method and Straw method for cryopreservation of human low volume sperm.Methods:A prospective study was performed to analyze the post-thaw results of cryopreserved human sperm using the home-made Strawtop method and Straw method. Compare the survival rate, sperm DNA fragmentation index (DFI), ultrastructure, fertilization ability and embryo developmental competence of donated semen samples from the patients who underwent intracytoplasmic sperm injection (ICSI) treatment in Center for Reproductive Medicine, Shanghai East Hospital, Tongji University School of Medicine during the period from May 1, 2018 to June 30, 2018.Results:The survival rate of Strawtop group (46.8%±17.1%) was significantly higher than that of Straw group (23.1%±13.7%, P=0.001). Compared with fresh sperm (18.9%±11.6%), Straw method (33.5%±15.0%) significantly increased the sperm DFI ( P=0.019), while Strawtop method didn't (23.4%±11.7%, P=0.375). The sperm ultrastructure of Strawtop group was superior to that of Straw group. There was no difference among Strawtop, Straw and fresh group in normal fertilization rates, cleavage rates and day 2 high-quality embryo rates. Conclusion:For human low volume sperm cryopreservation, the Strawtop method was simple and convenient. The survival rate of Strawtop method was higher than that of straw method and the sperm could be used for ICSI directly after thawing. The strawtop method is worth to be widely used in clinical practice.

Objective:To evaluate the cryopreservation effect of the home-made Strawtop micro-carrier method and Straw method for cryopreservation of human low volume sperm.Methods:A prospective study was performed to analyze the post-thaw results of cryopreserved human sperm using the home-made Strawtop method and Straw method. Compare the survival rate, sperm DNA fragmentation index (DFI), ultrastructure, fertilization ability and embryo developmental competence of donated semen samples from the patients who underwent intracytoplasmic sperm injection (ICSI) treatment in Center for Reproductive Medicine, Shanghai East Hospital, Tongji University School of Medicine during the period from May 1, 2018 to June 30, 2018.Results:The survival rate of Strawtop group (46.8%±17.1%) was significantly higher than that of Straw group (23.1%±13.7%, P=0.001). Compared with fresh sperm (18.9%±11.6%), Straw method (33.5%±15.0%) significantly increased the sperm DFI ( P=0.019), while Strawtop method didn't (23.4%±11.7%, P=0.375). The sperm ultrastructure of Strawtop group was superior to that of Straw group. There was no difference among Strawtop, Straw and fresh group in normal fertilization rates, cleavage rates and day 2 high-quality embryo rates. Conclusion:For human low volume sperm cryopreservation, the Strawtop method was simple and convenient. The survival rate of Strawtop method was higher than that of straw method and the sperm could be used for ICSI directly after thawing. The strawtop method is worth to be widely used in clinical practice.

[Objective]To evaluate the role of mature cumulus cells from oocyte-cumulus complex(OCC)in in-vitro maturation(IVM)and establish a new culture technique which is convenient to carry out.[Methods]The cumulus cells of OCC were cut off and dispersed by 1 mL syringe.The cumulus cells were co-cultured with the immature oocytes retrieved from the COH cycles after they adherent to the bottom of the dish.The immature oocytes were experienced IVM procedures in different culture media.They were divided into 3 groups(the oocytes at germinal vesicle stage from one woman were allotted to the same group randomly).Group 1(solution A):basic culture medium+human follicular fluid(hFF);Group 2(solution B):solution A+ cumulus cells(OCC);Group 3(solution C):solution A+ OCC+ follicle stimulating hormone(FSH)+ epidermal growth factor (EGF).Then,the maturation rate,fertilization rate and formation rate of available embryo were observed.[Results]In 113 treatment cycles,298 immature oocytes were performed IVM with solution A,B,and C.The difference for 24 hour maturation rates among 3 groups wag statistically significant(A:45.2%,B:61.7%,C:78.2%,P＜0.05).There was no statistical difference for 25～48 hour maturation rates and normal fertilization rates of mature oocytes.The differences of cleavage rates and rescued embryo rates between group 1 and 2,group 1 and 3 were statistically significant(P＜0.05).The formation rates of available embryo showed an increasing trend from group 1,2,to 3.[Conclusion]After being dispersed by simply beat upon with syringe and adherent culture,the mature cumulus eells from mature OCCs in COH cycles,together with growth factors in the follicular fluid or extraneously supplemented,could promote the IVM of immature oocyte.