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Establishment and optimization of the culture system of mouse oocyte in vitro maturation
GE Li DU Hui LIU Li-wei SU Yan-ping*
Acta Anatomica Sinica ›› 2013, Vol. 44 ›› Issue (4) : 535-540.
Establishment and optimization of the culture system of mouse oocyte in vitro maturation
Objective To study the effects of the duration of pregnant mare serum gonadotropin(PMSG) priming and in vitro culture (IVC) on nuclear maturation, and the effects of the different activation schemes on the abilities of parthenogenetic activation
and development. Methods To detect the rate of nuclear maturation at different time points of IVC by using PMSG priming(each treatment had at least 3 replicates, 3 mice per replicate).In order to determine the optimum activation scheme, two schemes
including: (1) ethanol combining with 6-dimethylaminopurine(6-DMAP) and (2) SrCl2 were used, CZB［fetal bovine serum(FBS) and CZB［bovine serum albumin(BSA)］ were used for embryo culture. In order to determine the optimum activation age, the oocytes matured
at the different time points were activated. To investigate the effect of PMSG on the ability of oocyte development by shortening the duration of PMSG priming. Results The time of IVC when oocytes reached the highest rate of nuclear maturation (97.6% vs 91.9%)
was prolonged from 14 hours to 16 hours if shortening the duration of PMSG priming from 46 hours to 24 hours. Shortening the duration of PMSG priming did not affect the rate of nuclear maturation, however, the rates of activation（91.2% vs 37.1%）and
blastocyst（20.9% vs 0.0%）were significantly reduced. Two schemes used in the present study were able to induce the activation rate of oocytes to more than 90% in the in vitro maturation system, however, the rates of blastocyst were significantly different
(P<0.05). The rates of activation (89.5%) and blastocyst (21.9%) reached the highest points from 24 hours to 26 hours during IVC. Conclusion A relatively ideal IVC system has been established in the present study. PMSG stimulation duration is 46 hours,
oocytes cultured for 24 hours, activated with CZB (10mmol/L SrCl2) and embryos cultured in CZB (0.5% BSA).
Oocyte / Parthenogenetic activation / In vitro culture / Embryo culture / Mouse
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