猪体外受精胚胎、孤雌激活胚胎以及体细胞核移植胚胎的体外培养

Preimplantational Development of Porcine Embryos Derived from in vitro Fertilization,Parthenogenetic Activation and Somatic Cell Nuclear Transfer

目的本研究系统比较了体外受精胚(IVFEs)、孤雌激活胚(PAEs)以及体细胞核移植胚(NTEs)的发育率和囊胚质量,同时还比较了PZM-3和NCSU-23两种培养基培养对PAEs以及NTEs发育的影响。方法利用体细胞核移植技术、体外受精技术和孤雌激活技术分别生产猪的NTEs、IVFEs和PAEs,开展体外培养。结果(1)IVFEs、PAEs和NTEs的卵裂率分别为72.0%,66.0%和63.5%,各组间没有显著差异(P>0.05);囊胚形成率分别为11.0%,22.6%和16.9%,也不存在明显不同(P>0.05);然而,在囊胚质量,即ICM细胞数/总细胞数的比例上,IVFEs和NTEs均优于PAEs(0.448%,0.356%vs 0.122%,P<0.05)。(2)对PAEs来讲,以PZM-3为基础培养基时囊胚率明显高于以NCSU-23为基础培养基时的处理组(35.4%vs.22.6%,P<0.05);而对NTEs而言,两种培养基在支持胚胎形成囊胚方面的效果相当(26.6%vs.21.1%,P>0.05),虽然PZM-3培养时胚胎卵裂率显著低于NCSU-23培养组(47.5%vs.63.5%,P<0.05)。结论(1)PAEs的囊胚质量在3种来源的胚胎中最差;(2)对PAEs理想的培养基,当用于NTEs培养时却不一定是最佳的选择。

[Objective] The present study were designed to examine the developmental competence of preimplantational embryos derived from in vitro fertilization, parthenogenetic activation and somatic cell nuclear transfer in pig, and to evaluate the quality of porcine blastocysts derived from IVF, PA and SCNT. In addition, the effects of porcine zygote medium 3 （PZM-3） and North Carolina State University Medium 23 （NCSU-23） on the developmental potential on the porcine PA embryos （PAEs） and SCNT （NTEs） embryos was also investigated. [Methods] NTEs, IVFEs and PAEs were produced by using somatic cell nuclear transfer, in vitro fertilization and parthenogenetic activation methods, followed by in vitro culture until use. [Results] （1） for the cleavage rate, no significant difference was observed among 1VF embryos （IVFEs）, PAEs and NTEs （72.0%, 66.0% and 63.5%, P〉0.05）, meanwhile for the blastocyst formation rate, no significant difference was obtained neither （11.0%, 22.6% and 16.9%, P〉0.05）. However, for the quality of blastocysts, i.e. the ratio of the cell number of inner cell mass （ICM） to total cell number, the IVFEs and NTEs were both superior to PAEs（0.448%, 0.356% vs. 0.122%, P〈0.05）. （2） as for PAEs, the blastocyst rate was significantly higher when cultured in PZM-3 than cultured in NCSU-23 （35.4% vs. 22.6%, P〈0.05） ; while as for NTEs, similar results of PZM-3 and NCSU-3 in supporting blastocyst formation were observed （26.6% vs. 21.1%, P〉0.05 ）, although the cleavage rate was obviously higher in PZM-3 group than NCSU-23 group （47.5% vs. 63.5%, P〈0.05）. [Conclusion] （1） the poorest quality of blastcysts was generally observed in PAEs; （2） the ideal culture medium for PAEs may not be necessarily beneficial to NTEs.