3种冷冻方法对猪MII期卵母细胞冷冻后微丝分布和脂肪颗粒变化的影响

doi:10.11843/j.issn.0366-6964.2014.07.010

畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (7): 1097-1103.doi: 10.11843/j.issn.0366-6964.2014.07.010

3种冷冻方法对猪MII期卵母细胞冷冻后微丝分布和脂肪颗粒变化的影响

吴彩凤^1,2，戴建军^1,2，张树山^1,2，李维杰³，徐利^1,2，张德福^1,2*

(1.上海市农业科学院畜牧兽医研究所，上海 201106； 2.上海农业遗传育种重点实验室动物遗传工程研究室，上海 201106；3.上海理工大学生物系统热科学研究所，上海 200093）

收稿日期:2014-01-25 出版日期:2014-07-23 发布日期:2014-07-23
通讯作者: 张德福，研究员，E-mail：zhangdefuzdf@163.com
作者简介:吴彩凤 (1983-)，女，黑龙江佳木斯人，硕士，助理研究员，主要从事动物胚胎工程研究，Tel：021-52235475，E-mail：wucaifengwcf@163.com
基金资助:
上海市科委农业成果转化专项（123919N0700;133919N1700）；国家自然科学基金（31372315）；高繁殖力转基因猪新品种培育（2013ZX08006-005）

Effects of Three Vitrification Methods on the Microfilament Distribution and Lipid Droplets Changes of Porcine MII-stage Oocytes

WU Cai-feng^1,2，DAI Jian-jun^1,2，ZHANG Shu-shan^1,2，LI Wei-jie³，XU Li^1,2，ZHANG De-fu^1,2*

(1.Institute of Animal Husbandry and Veterinary Science，Shanghai Academy of Agricultural Sciences，Shanghai 201106，China； 2.The Research Group of Animal Genetic Engineering，Shanghai Municipal Key Laboratory of Agri-genetics and Breeding，Shanghai 201106，China； 3.Institute of Biothermal Science and Technology，University of Shanghai for Science and Technology,Shanghai 200093，China)

Received:2014-01-25 Online:2014-07-23 Published:2014-07-23

摘要/Abstract

摘要：

本研究旨在探讨3种冷冻方法（CLV法、OPS法和Straw法）对猪MII期卵母细胞玻璃化冷冻后微丝、脂肪颗粒分布及其超微结构的影响。结果显示：（1）CLV(Cryoloop vitrification)法冷冻速率最高，达42 639 ℃•min^-1；（2）CLV法微丝正常分布率（47.06%）最高，显著高于OPS(Open pulled straw)法的38.04%和Straw法的30.95%（P<0.05）。（3）CLV法和OPS法的脂肪颗粒正常分布率之间没有显著差异(P>0.05)，但两者均显著高于Straw法(P<0.05)；（4）解冻后卵母细胞超微结构观察结果显示，大的脂肪颗粒含量明显减少；大部分脂肪颗粒冷冻后呈均质化状态，只有很少的异质化脂肪颗粒存在。试验结果表明，冷冻过程中卵母细胞脂肪颗粒破坏严重，增加冷冻速率能降低对微丝的损伤，从而减少对脂肪颗粒异常分布的影响。

Abstract:

The purpose of this study was to determine the microfilament distribution，lipid droplets and their ultrastructure changes that resulted from vitrified porcine MII-stage oocytes with 3 vitrification techniques（Straw method，CLV method and OPS method）.The results showed that：(1) The cooling speed of CLV method was the highest，and got 42 639 ℃•min^-1.(2) CLV method got the highest normal distribution rate (47.06%)，and it was much higher than OPS method (38.04%) and Straw method (30.95%).(3) There was no significant differences in the normal distribution rate of lipid droplets between CLV and OPS methods after thaw，but they were much higher than that of Straw method (P<0.05).(4) Decreased content of large lipid droplets in thawed oocytes were found out after ultrastructural observation with transmission electron microscopy (TEM)，and most of lipid droplets in thawed oocytes were homogeneous and very few of them were heterogeneous.These results concluded that the lipid droplets and their distribution after vitrification were damaged greatly.Improving cooling speed could decrease the injury of microfilament，and then reduce the damage degree of distribution in lipid droplets.

中图分类号:

S814.8

吴彩凤，戴建军，张树山，李维杰，徐利，张德福. 3种冷冻方法对猪MII期卵母细胞冷冻后微丝分布和脂肪颗粒变化的影响[J]. 畜牧兽医学报, 2014, 45(7): 1097-1103.

WU Cai-feng，DAI Jian-jun，ZHANG Shu-shan，LI Wei-jie，XU Li，ZHANG De-fu. Effects of Three Vitrification Methods on the Microfilament Distribution and Lipid Droplets Changes of Porcine MII-stage Oocytes[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(7): 1097-1103.

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3种冷冻方法对猪MII期卵母细胞冷冻后微丝分布和脂肪颗粒变化的影响

Effects of Three Vitrification Methods on the Microfilament Distribution and Lipid Droplets Changes of Porcine MII-stage Oocytes

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