摘要
猪卵母细胞在TCM—199培养基中,20℃和37℃(对照)下,分别培养24h和48h。培养基中加入20%混合的3个犊牛血清、5%的猪卵泡液。气体浓度为90%N_2,5.2%O_2和4.8%CO_2。24h,20℃和37℃培养的卵母细胞经台盘蓝鉴别后,存活率分别为71%和69%。当培养到48h,20℃组的成活率明显高于37℃组,分别为61%和25%。电镜观察,20℃组卵母细胞的线粒体较完整,微绒毛较多。而37℃组则相反。当20℃培养24h 后,把卵母细胞移到37℃下继续培养24h,仍有45%的存活率,11%的颗粒细胞扩散程度较好。染色后,可见第1极体和染色体,说明处于第2次成熟分裂的中期。实验说明,20℃保存24h 仍有继续发育成熟的潜力。
The porcine oocytes were cultured in TCM-199 at 20℃ and 37℃(control)for 24h and48h,respectively.The culture medium TCM-199 contained 20% calf serum from three calvesand 5% porcine follicular fluid.The gas concentrations were 90% N_2,5.2% O_2 and 4.8%CO_2.The survival rates of oocytes under 20℃ and 37℃ for 24 h were 71% and 69%respectively as judged through staining with trypan blue.The survival rate of oocytes culturedat 20℃ for 48h(61%)was obviously higher than that cultured at 37℃(25%).The result ofobservation with an electronic microscope was that the mitochondria in 20℃ group had acomplete structure and there were more microvilli on the oocytes in this group.After culturedat 20℃ for 24h,the oocytes were cultured at 37℃ for 24 h and their survival rate was 45%,with 11% cumuli spreading well.After being stained,the first polar bodies and chromosomescould be seen,which showed that the oocytes were at the second metaphase.The experimentshowed that the porcine oocytes still have continuously developmental and maturationcapecity under normal conditions after 24 h preservation at 20℃.
关键词
猪
卵母细胞
保存
培养
pig
porcine oocyte
preservation at room temperature
maturation in culture
