| Objective:To evaluate the effect of MSC-EVs on reconstruction of mouse ovarian function and fertility.Methods:All experimental mice were divided into wide type group?WT?,premature ovarian failure group?POF?and EVs treatment group?EVs?,with 40 mice for each group.The POF model was established through intraperitoneal injection of cyclophosphamide?200 mg/kg?and busulfan?20 mg/kg?.Two weeks later,0.1 ml 150?g MSC-EVs were injected through tail vein in EVs group,while equal volumes of PBS were injected in WT group and POF group.General condition and body weight of each group were observed regularly.4 weeks after MSC-EVs injection,8 mice were sacrificed for evaluation of ovarian function through serous level of E2 and FSH,histology and immunohistochemistry of ovary.Apoptosis-related mRNA and protein expression level including Capase-3,Bcl-2 and Bax were detected by qPCR and western blot.6 mice were performed ovarian stimulation for evaluation of the outcome of IVF,and 15 were chosen for mating test.The offspring of each group received Y-maze test and novel object recognition test for evaluation of their learning and memory ability.Results:?1?14 days after CTX/BUS injection,the body and ovarian weight decreased significantly and vaginal smear showed irregular oestrous cycle.The level of E2decreased and FSH increased significantly.Histology and immunohistochemistry showed a significant reduction of follicles in every stage and up-expression of Capase-3.Open field test indicated that the locomotor activity of mice weakened following increased anxiety.In process of IVF,the number of oocyte retrieved,fertilized embryos,cleaved embryos and blastocysts in POF group were all significantly deceased compared with WT group.Mating test showed no offspring obtained in POF group.?2?4 weeks after MSC-EVs injection,the body and ovary weight in EVs group were significantly higher than those in POF group,with the level of E2 increased and FSH decreased.Histology and immunohistochemistry of ovary showed that the number of primordial follicles,primary follicles and secondary follicles all increased in EVs group.Furthermore,the outcomes of IVF such as oocyte retrieved,fertilized oocytes,cleaved embryos and blastocysts in EVs group were all increased compared with POF group.Mating test showed 11 out of 15 female mice in EVs group delivered,4 out of 15female mice in POF group delivered,all the mice in WT group delivered.The apoptosis associated genes?Caspase 3,Bcl-2?and its protein expression decreased in EVs group compared with group POF based on qPCR and western blot analysis.?3?The body weight of offsprings increased gradually,while WT group showed heavier than the other two groups at the first week after birth,then no significant difference were identifited in offspring of each group until 6 weeks after birth.Y-maze test and novel object recognition test in offspring of each group showed no significant diffenence.Conclusion:?1?MSC-EVs could reconstruct and recover the ovarian function and fertility of POF mice induced by CTX/BUS.?2?MSC-EVs could inhibit the apoptosis of granulosa cell induced by CTX/BUS.?3?The learning and memory ability of offspring were not affected by MSC-EVs treatment.Objective: To evaluate the effect of optimal MSC-EVs administration on fertility reconstruction of premature ovarian failure mouse.Methods: All experimental mice were divided into wide type group?WT?,premature ovarian failure group?POF?and EVs treatment group?EVs?,with 40 mice for each group.The POF models were established through intraperitoneal injection of cyclophosphamide?200 mg/kg?and busulfan?20 mg/kg?.Two weeks later,EVs group was subdivided into once administration group?once-EVs?and multiple administration group?multi-EVs?,then 0.1 ml 150 ?g MSC-EVs were injected through tail vein once in once-EVs group and once a week for 4 times in multi-EVs group.Meanwhile,equal volumes of PBS were injected in WT group and POF group respectively.Then some mice were sacrificed 1,2,3 and 4 weeks after MSC-EVs administration respectively for evaluation of ovarian function through serous level of E2 and FSH,histology and immunohistochemistry of ovarian.The others received IVF and mating test for evaluation of fertility of the mice.Results:?1?The effects of once MSC-EVs administration on fertility reconstruction: The body and ovary weight in once-EVs group recovered gradually,and were significantly higher than POF group.The serum level of E2 increased and FSH decreased significantly in once-EVs group compared with POF group 2 weeks later,while no significant difference 4 weeks later.Histology of ovary showed that the numbers of primordial follicles,secondary follicles and antral follicles in once-EVs group all increased significantly 3 weeks later.?2?Comparison between once and multiple MSC-EVs administration: The outcomes of IVF and mating test showed there were no significant difference between once-EVs group and multi-EVs group.Conclusion: Both once and multiple MSC-EVs administration could recover the ovary function and fertility of POF mice. |
PDF Full Text Request