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绵羊卵母细胞体外成熟过程中大分子来源和培养基类型对后续胚胎发育的影响

The Effect of Macromolecule Source and Type of Media During in vitro Maturation of Sheep Oocytes on Subsequent Embryo Development.

作者信息

Shirazi Abolfazl, Ardali Mohammad Ansari, Ahmadi Ebrahim, Nazari Hassan, Mamuee Morteza, Heidari Banafsheh

机构信息

Department of Embryology and Andrology, Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran ; Department of Gametes and Cloning, Research Institute of Animal Embryo Technology, Shahrekord University, Shahrekord, Iran.

出版信息

J Reprod Infertil. 2012 Jan;13(1):13-9.

Abstract

BACKGROUND

Oocyte maturation and subsequent in vitro production (IVP) of embryos are affected by diverse groups of chemicals in maturation medium which are needed for successful mammalian oocyte maturation during which the dramatic cytoplasmic and nuclear reprogramming events take place. This study was designed to evaluate the effects of protein source (fetal bovine serum, FBS, and bovine serum albumin, BSA) as well as two different maturation media during in vitro maturation of ovine oocytes on subsequent embryo development.

METHODS

Cumulus oocyte complexes were recovered from ovaries obtained from slaughter house and cultured for 24 hr in either TCM-199 or SOFaa maturation medium supplemented with 10% (v/v) FBS or 0.8% (w/v) BSA. Data were analyzed by one-way ANOVA using Sigma Stat (Ver. 2). A p-value smaller than 0.05 was considered statistically significant.

RESULTS

The proportions of cleavage and total blastocyst (evaluated on days 3 and 6, respectively) were significantly higher in FBS than BSA supplemented groups, though no differences were observed between the two used different maturation media. The cryotolerance of blastocysts was negatively influenced by the presence of FBS rather than BSA during IVM. The quality of produced embryos, however, was affected neither by the source of macromolecules nor the maturation medium in terms of hatching rate, total blastocyst cells and inner cell mass/total cell ratio.

CONCLUSION

The rate of oocyte development was improved by the presence of FBS, though the cryosurvival of resulting blastocysts was negatively influenced by the presence of the serum during in vitro production of sheep oocytes.

摘要

背景

卵母细胞成熟及随后的胚胎体外生产(IVP)会受到成熟培养基中多种化学物质的影响,而这些化学物质是哺乳动物卵母细胞成功成熟所必需的,在此过程中会发生显著的细胞质和细胞核重编程事件。本研究旨在评估蛋白质来源(胎牛血清,FBS,和牛血清白蛋白,BSA)以及两种不同的成熟培养基在绵羊卵母细胞体外成熟过程中对后续胚胎发育的影响。

方法

从屠宰场获取的卵巢中回收卵丘卵母细胞复合体,并在补充有10%(v/v)FBS或0.8%(w/v)BSA的TCM - 199或SOFaa成熟培养基中培养24小时。使用Sigma Stat(版本2)通过单因素方差分析对数据进行分析。p值小于0.05被认为具有统计学意义。

结果

在第3天和第6天分别评估时,FBS补充组的卵裂率和总囊胚率显著高于BSA补充组,尽管在两种使用的不同成熟培养基之间未观察到差异。在体外成熟过程中,FBS的存在对囊胚的耐冻性有负面影响,而不是BSA。然而,就孵化率、总囊胚细胞数和内细胞团/总细胞比率而言,所产生胚胎的质量既不受大分子来源的影响,也不受成熟培养基的影响。

结论

FBS的存在提高了卵母细胞的发育率,尽管在绵羊卵母细胞体外生产过程中血清的存在对所得囊胚的冷冻存活率有负面影响。

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Role of protein supplements in the culture of mouse embryos.
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