| Objective:To explore the effects of rat serum containing Youguiwan (YGW) on the function of programmed cryopreservation of mouse ovarian tissue after recovery.Methods:Cryopreserved ovaries were obtained from406-week-old ICR mice which were stimulated by Pregnant Mare Serum Gonadotropin (PMSG). All ovarian tissue were cryopreserved in liquid nitrogen by programmed cryopreservation. and recovered after2days. Then all ovaries were randomly assigned into4group, including blank group, blank serum group. FSH group and YGW serum group. Each ovarian tissue was placed with DMEM medium into four groups, which included none sereum,3%v/v8#rat serum (blank serum),0.3IU/ml FSH and3%v/v YGW rat serum respectively, and each group contained20ovaries.24h later, the immature oocytes were extracted from left ovaries, and cultivated by micro-drip method for another24h. Finally, the first polar body of oocytes in vitro (PB1) exclusion rate was measured via micro-drips under a microscope and inverted microscope, and the content of E2in the medium, and the content of MPF, FSH-R and LH-R in the right ovaries were calculated by enzyme-linked immunosorbent assay (ELISA). All the data was analyzed by SPSS19.0software.Results:1After recovery, the oocytes maturation rate of YGW serum group was significantly higher than that of FSH positive control group and blank serum group (P<0.05).2After recovery, the level of MPF and FSH-R of YGW group were significantly higher than that of FSH positive control group and blank group (P<0.01). The blank serum group had higher levels of MPF and FSH-R as well, which was significantly higher than that of FSH positive control group and blank group (P<0.01). However, there was no significant difference between YGW serum group and blank group (P>0.05).3After recovery, the level of LH-R in YGW serum group was significantly higher than that of FSH positive control group and blank group (P<0.01). and was also higher than that of blank serum group (P<0.05). The level of LH-R of blank serum group was higher than that of blank group (P<0.05).4After recovery and in vitro culture for24h, E2concentration in the culture medium in YGW serum group was higher than that of blank serum group (P<0.05), and significantly higher than blank group (P<0.01). E2concentration in the culture medium, FSH group and blank serum were significantly higher than those of blank group (P<0.01).Conclusions:1YGW rat serum can improve the function of ovarian tissue after recovering from programmed cryopreservation.2YGW rat serum might improve the level of maturation promoting factor in mice (MPF). follicle-stimulating hormone receptor (FSH-R) and luteinizing hormone receptor (LH-R) to get a better function of ovarian tissue after being recovered from programmed cryopreservation.3The increase of the level of luteinizing hormone receptor (LH-R) of oocytes after programmed cryopreservation and recovery induced by YGW rat serum, might be achieved by raising LH-R expression level directly as well as follicular FSH-R indirectly to improve LH-R expression level.4YGW rat serum is the effective means of promoting in-vitro maturation of oocytes in ovarian tissue after programmed cryopreservation and recovery.5Drug interventions after recovering from programmed cryopreservation is one of the effective interventions. |
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