猪体外成熟卵母细胞的电激活及其激活后的体外培养  

Studies on Electrical Activation of Porcine Oocytes Matured in vitro and Embryo Culture Systems

作　　者：吴中红[1] 邢凤英[1] 刘国世[1] 曾申明[1] 朱士恩[1] 张忠诚[1] 伏彭辉 

机构地区：[1]中国农业大学动物科技学院,北京100094

出　　处：《中国农业科学》2002年第12期1537-1542,共6页Scientia Agricultura Sinica

基　　金：国家自然科学基金 (3 0 10 0 13 3 );香港WISS生物技术研究发展中心资助项目

摘　　要：主要研究了猪体外成熟卵母细胞电激活的条件以及孤雌激活胚胎的体外培养体系。用不同场强、脉冲时程和脉冲次数激活猪体外成熟的卵母细胞。结果表明 ,在本试验条件下电激活最佳场强和脉冲时程为 130V·mm-1/ 80 μs ,其囊胚发育率为 (2 0 .12± 8.18) % (P >0 .0 5 )。多次脉冲并不比单次脉冲的激活效果好 (P >0 .0 5 )。孤雌激活胚胎用不同培养方式和气体环境在体外培养 7d ,发现 7d不换液与第 5天换液和第 5天换含 10 %胎牛血清的培养液其囊胚发育率 [分别为 (2 5 .30± 7.5 5 ) %、(2 6 .4 4± 8.35 ) %和 (17.6 8± 5 .39) % ]无显著差异 (P >0 .0 5 ) ,后两者换液培养方式其囊胚细胞数 (15 .78± 5 .4 6、14 .5 5± 4 .81)明显低于不换液的 (18.0 1± 6 .79,P <0 .0 1)。同时发现低氧 (5 %CO2 ∶7%O2 ∶88%N2 )气体环境培养的胚胎囊胚发育率和囊胚细胞数和高氧环境 (5 %CO2 的空气 )无明显差别 ,分别为 [(2 0 .78± 8.80 ) %、17.0 0± 6 .12和 (2 5 .30± 7.5 5 ) %、18.0 1± 6 .79,P >0 .0 5 ]。表明激活后第 5天换原液和换含胎牛血清的培养液不能提高囊胚细胞数。低氧 (5 %CO2 ∶7%O2 ∶88%N2 )气体环境培养猪胚胎的效果不比高氧环境 (5 %CO2 的空气 )好。Conditions for electrical parthenogenetic activation of porcine oocytes matured in vitro and in vitro culture systems of porcine embryo were studied. The best results were achieved under the conditions of electrical field strength and the pulse duration at 130V·mm -1 /80μs, with a blastocyst development rate of (20.12± 8.18 )% (P>0.05). No significant deference was found between treatments of multiple pulses and a single pulse (P>0.05). Parthenogenetic embryos were cultured with different methods and air conditions for 7 days in vitro, blastocyst development rate of embryos with changed culture media [(26.44±8.35)%] or changed media with 10% fetal bovine serum (FBS) [(17.68±5.39)%] on the fifth day showing no significant diffe rence from that of embryos without change of culture media [(25.30±7.55)%, [WTBX]P>0.05], while the cell numbers of blastocysts from embryos with changed culture media (15.78±5.46 and 14.55±4.81) were significan tly lower than the number of blastocysts from embryos without change of culture media (18.01±6.79, , while the cell numbers of blastocysts from embryos with changed culture media (15.78±5.46 and 14.55±4.81) were significan tly lower than the number of blastocysts from embryos without change of culture media (18.01±6.79, P<0.01). Blastocyst development rate and blastocyst cell number of embryos cultured in lower O 2 (5%CO 2∶7%O 2∶88%N 2) also showed no significant difference from those in high O 2 (5%CO 2 in air) [(20.78±8.80)% and 17.00 ±6.12 vs. (25.30±7.55)% and 18.01±6.79, [WTBX]P>0.05]. It is concluded that change of culture media with the same new one or changing over to media with 10% fetal bovine serum (FBS) on the fifth day and low O 2 environment are not necessary for porcine embryos development. . It is concluded that change of culture media with the same new one or changing over to media with 10% fetal bovine serum (FBS) on the fifth day and low O 2 environment are not necessary for porcine embryos development.

关 键 词：猪 体外成熟卵母细胞 电激活 体外培养 胚胎培养 

分 类 号：S828[农业科学—畜牧学]