Study Of Allograft Vitrification Cryopreserved Newborn Mice Ovarian Tissue In Male Recipient Mice

The cryopreservation and transplantation of ovarian tissue is being considered one of new technique to build mammal ovum library in the future. To exploring methods of vitrification of ovarian tissue and the ideal cryopretectants and building the cryopreservation and transplantation technology of livestock ovary tissue, the vitrified newborn mice ovarian tissue were allografted underneath the kidney capsule of male recipients to in the experiment.In the study,the use of EG (ethylene glycol) and DAP (Dimethyl sulfoxidev,Acetamide and Propylene glycol) as cryoprotectants,1-day-old mice ovarian tissue were theated by two kinds of difference cryoprotectants and two kinds difference vitrification methods, dividaturing four groups(the cryovial method vitrification method of DAP,the direct cover vitrification method of DAP,the cryovial method vitrification method of EG and the direct cover vitrification method of EG). The thawing ovarian tissue were grafted the kidney capsule of male mice recepients, Oarian grafts were recovered 21-28 days after transplantation. Development of the oocytes and follicles of ovarian grafts and expression of genes relate to follicular development were evaluated with morphology,histology and semiquantitative RT-PCR.1.one hundred and two male recipient used and two hundred and seven ovaries form 1-day-old mice grafted; the grafts were successfully recovered form seventy two of the male recipients and the recovery rate was 70.5% in total; the ovary were successfully recovered form one hundred of the male recipients and thirteen and the recovery rate was 54.5% in total.all Ovary grafts were very significantly grew(P< 0.01) , the grafts were best grew in the 28d-ET group.2.Primordial follicle formation,recruitment and growth can be initiated in cryopreserved ovarian grafts in male recipient mice;Ovarian grafts contained gollicles at all stages of folliculogenesis but didn't find corpus luteum on 21-28 days after transplantation.3.Oocytes collected from the grafts and the yield of oocytes per ovarian graft in the 21dDT, 28dDT,21d-ET and 28d-ET groups were 12.6±7.3,8.8±8.6,10.1±2.4 and 35.0±50.3,respectively.the difference is not significant(P>0.05). 134 and 55 oocytes were respectively collected from two ovary in the 28d-ET group.4.BAX and P450—arom were detected in normal ovaries,ovarian grafts and thewing ovaries.Expression of target gens relate to follicular development and follicle atresia was not influenced by vitrification and transplantation.in conclusion,the pattern of follicular development and follicle atresia in the thewing ovary grafts may be similar to the ovaries of normal female mice.The follicles in ovarian grafts from intact male recipients can grow and develop to maturation stage.The cryopreserved ovarian tissue can well be preserved; The thawing 1-day-old mice ovarian tissue can grow and develop after allografting;The grafts can gain many oocytes by separation.