亮甲酚蓝对猪卵母细胞体外成熟及核移植胚胎发育能力的影响

doi:10.16431/j.cnki.1671-7236.2016.03.028

Abstract

Abstract: This experiment aimed to study the effect of brilliant cresyl blue (BCB) on in vitro maturation of pig oocytes and the developmental capacity of pig SCNT embryos.The cumulus-oocyte complexes (COCs) were stained with different concentrations of BCB (13,26,39 and 52 μmol/L) for 90 min,and then we divided the COCs into BCB⁺ and BCB^- for in vitro culture 42 to 44 h.The results showed that,with the concentration of BCB increased,the staining rate (20.00%,46.39%,51.66% and 59.03%) raised gradually while the maturation rate of oocytes (74.03%,72.16%,70.53% and 48.61%) reduced,the percentages of oocytes staining by 26 μmol/L BCB for 90 min were higher than that of other groups in staining rate and maturation rate.However,the nuclear maturation rate of BCB⁺ groups were higher than that of BCB^- group.Therefore,26 μmol/L BCB was selected as the most effective concentration dying the oocytes (BCB⁺),which were used as parthenogenetic activation and nuclear transfer embryos.The cleavage and blastocyst rates of parthenogenetic activation and SCNT embryos in BCB⁺ group were significantly higher than that of BCB^- group (P<0.05),but there were no significant differences between the cleavage and blastocyst rates in the groups of BCB⁺ and control (P>0.05).Reconstructed embryos derived from the COCs stained with BCB were transferred to five surrogates,and six cloned piglets were obtained from one of the two pregnant pigs.These results showed that COCs stained with BCB was an effective method to select high-quality oocytes,which could improve the efficiency of in vitro embryo production.

Key words: pig; oocyte; in vitro maturation (IVM); brilliant cresyl blue (BCB); parthenogenetic activation; nuclear transfer

CLC Number:

S814.8

XU Guo-qi, HUA Zai-dong, XU Sheng-cheng, ZHENG Xin-min. Effect of BCB Staining on in vitro Maturation and Development of Nuclear Transfer Embryo of Pig Oocyte[J]. , 2016, 43(3): 754-760.

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Effect of BCB Staining on in vitro Maturation and Development of Nuclear Transfer Embryo of Pig Oocyte

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