不同因素对猪卵母细胞体外成熟培养的影响

›› 2014, Vol. 41 ›› Issue (10): 205-209.

不同因素对猪卵母细胞体外成熟培养的影响

林筱璐¹, 林秀娇¹, 王秀爱², 刘凤军³, 庄益芬¹, 张兴锋¹, 李鸿翔¹, 洪志勇¹, 刘芳¹, 张文昌¹

1. 福建农林大学动物科学学院, 福建福州 350002;
2. 福清市永诚畜牧有限公司, 福建福清 350319;
3. 河南科技大学动物科技学院, 河南洛阳 471003

收稿日期:2014-04-08 出版日期:2014-10-20 发布日期:2014-10-29
通讯作者: 张文昌 E-mail:zwcfz@163.com
作者简介:林筱璐(1990－),女,福建人,硕士生,研究方向:动物繁殖新技术;林秀娇(1990－),女,福建人,硕士生,研究方向:动物繁殖生理与胚胎工程。
基金资助:
福建省科技计划项目(2008N0001)

Effect of Different Factors on in Vitro Maturation of Porcine Oocytes

LIN Xiao-lu¹, LIN Xiu-jiao¹, WANG Xiu-ai², LIU Feng-jun³, ZHUANG Yi-fen¹, ZHANG Xing-feng¹, LI Hong-xiang¹, HONG Zhi-yong¹, LIU Fang¹, ZHANG Wen-chang¹

1. College of Animal Science, Fujian Agriculture and Forestry University, Fuzhou 350002, China;
2. Fuqing Yongcheng Livestock Co., Ltd., Fuqing 350319, China;
3. College of Animal Science and Technology, Henan University of Science and Technology, Luoyang 471003, China

Received:2014-04-08 Online:2014-10-20 Published:2014-10-29

摘要/Abstract

摘要： 本试验旨在探讨不同外源激素组合的添加、胰岛素—转铁蛋白—亚硒酸钠(ITS)的添加、不同培养皿、石蜡油的添加对卵母细胞体外成熟培养(IVM)的影响。结果显示:①孕马血清促性腺激素+人绒毛膜促性腺激素+促卵泡素(PMSG+HCG+FSH)组高于促卵泡素+促黄体素(FSH+LH)组和尿促性腺激素(hMG)组,相互之间差异显著(80.1%、68.3%、53.3%,P<0.05);②添加1% ITS到猪卵母细胞培养液中对卵母细胞成熟率无显著提高(P>0.05),但显著提高了孤雌激活后孤雌胚胎的卵裂率和囊胚率(63.3%、55.1%,18.7%、12.1%,P<0.05);③凹槽皿组猪卵母细胞成熟率显著高于四孔板和30 mm塑料皿组(73.3%、68.0%、68.3%,P<0.05);④石蜡油的添加对卵丘细胞扩展和卵母细胞体外成熟率均有显著提高(84.8%、69.9%,P<0.05)。结果表明培养液中添加PMSG+HCG+FSH和ITS及选用凹槽皿、成熟培养液上覆石蜡油可提高猪卵母细胞IVM效果。

关键词: 猪卵母细胞; 体外成熟培养(IVM); 培养方式; 石蜡油; 外源激素; 胰岛素—转铁蛋白—亚硒酸钠(ITS)

Abstract: Effects of several factors such as exogenous hormones and ITS,culture methods,paraffin oil on in vitro maturation (IVM) for porcine oocytes were studied.The results indicated that,the group supplemented with PMSG+HCG+FSH was significantly higher than FSH+LH and HMG in mature media with regard to the mature rates of porcine oocytes (80.1% vs 68.3% vs 53.3%,P<0.05);the mature rate of porcine oocytes cultured in mature media supplemented with 1% ITS was not improved,while the cleavage rate and blastocyst rate of parthenogenesis embryos were significantly increased (63.3% vs 55.1%,18.7% vs 12.1%,P<0.05);the groove dish group was significantly higher than four-hole plate and 30 mm plastic dish group in terms of the mature rates of porcine oocytes (73.3% vs 68.0% vs 68.3%,P<0.05);mature media supplemented with paraffin oil was significantly increased the cumulus expansion and maturation rates for porcine oocytes (84.8% vs 69.9%,P<0.05). The results showed that the mature rates of porcine oocytes cultured in groove dish,supplemented with paraffin oil,PMSG+HCG+FSH and ITS were significantly improved.

Key words: porcine oocytes; in vitro maturation (IVM); culture methods; paraffin oil; exogenous hormones; ITS

中图分类号:

S828

林筱璐, 林秀娇, 王秀爱, 刘凤军, 庄益芬, 张兴锋, 李鸿翔, 洪志勇, 刘芳, 张文昌. 不同因素对猪卵母细胞体外成熟培养的影响[J]. , 2014, 41(10): 205-209.

LIN Xiao-lu, LIN Xiu-jiao, WANG Xiu-ai, LIU Feng-jun, ZHUANG Yi-fen, ZHANG Xing-feng, LI Hong-xiang, HONG Zhi-yong, LIU Fang, ZHANG Wen-chang. Effect of Different Factors on in Vitro Maturation of Porcine Oocytes[J]. , 2014, 41(10): 205-209.

参考文献

1 祁秀娟,陈振宇,简领,等.IVF实验室质控中培养液准备方法与早期鼠胚发育的关系[J].青岛医学院学报,1996,32(3):222~224.
2 黄雅琼,石德顺,张晓溪,等.猪卵母细胞体外成熟的发育潜力及核移植重构胚构建方法的研究[J].基因组学与应用生物学,2009,28(5):913~920.
3 杨喜,王志刚,刘丑生,等.猪卵母细胞的体外成熟影响因素的研究[J].中国畜牧兽医,2007,34(3):74~76.
4 Daniaux C,Verhaeghe B,Donnay I.Insulin,transferrin and selenium with or without BSA in a serum-free culture system for bovine embryo,and its suitability for embryos cultured in small groups[J].Reprod Fertil Dev,2005,17(2):217~218.
5 Hammer R E.Egg culture:The foundation[J].Int J Dev Biol,1998,42(7):833~839.
6 Jacobs S,Cuatrecasas P.Insulin receptor:Structure and function[J].Endocrine Rev,1981,2:251~263.
7 Machaty Z,Day B N,Prather R S.Development of early porcine embryos in vitro and in vivo[J].Biol Reproa,1998,59:451~455.
8 Markstrom E,Svensson E C,Shao R,et al.Survival factors regulating ovarian apoptosis-dependence on follicle differentiation[J].Reproduction,2002,123:23~30.
9 Nagai T.The improvement of in vitro maturation systems for bovine and porcine oocytes[J].Theriogenology,2001,55(6):1291~1301.
10 Onishi A,Iwamoto M,Akita T,et al.Pig cloning by microinjection of fetal fibroblast nuclei[J].Science,2000,289:1188~1190.
11 Palasz A T,Thundathil J,Verrall R E,et al.The effect of macromolecular supplementation on the surface tension of TCM-199 and the utilization of growth factors by bovine oocytes and embryos in culture[J].Anim Reprod Sci,2000,58:229~240.
12 Polejaeva I A,Chen S H,Vaught T D,et al.Cloned pigs produced by nuclear transfer from adult somatic cells[J].Nature,2000,407:86~90.
13 Raghu H M,Nandi S,Reddy S M. Effect of insulin,transferrin and selenium and epidermal growth factor on development of buffalo oocytes to the blastocyst stage in vitro in serum-free,semidefined media[J].Vet Rec,2002,151(9):260~265.
14 Shamsuddin M. A serum-free,cell-free culture system for development of bovine one-cell embryos up to blastocyst stage with improved viability[J].Theriogenology,1994,41:1033~1044.

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