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有血清培养与无血清培养的小鼠卵母细胞胚胎发育能力的比较。

Comparison of embryonic developmental competence of mouse oocytes grown with and without serum.

作者信息

Eppig J J, Wigglesowrth K, O'Brien M J

机构信息

Jackson Laboratory, Bar Harbor, Maine 04609.

出版信息

Mol Reprod Dev. 1992 May;32(1):33-40. doi: 10.1002/mrd.1080320107.

Abstract

The first objective of this study was to determine whether oocyte growth in serum-free medium affects the solubility of the zona pellucida to alpha-chymotrypsin digestion, which is an index of zona pellucida "hardening" and reflects the potential penetrability of the zona pellucida by sperm. Oocyte-granulosa cell complexes were isolated from the preantral follicles of 12-day-old mice and cultured for 10 days in medium containing 5% fetal bovine serum (FBS) or in serum-free medium. The zonae pellucidae of oocytes grown in serum-free medium were four times as hard as freshly isolated germinal vesicle (GV)-stage oocytes grown in vivo or oocytes grown in vitro in FBS-containing medium. The hardening of the zonae pellucidae of oocytes grown in serum-free medium was prevented by addition of fetuin. The second objective was to compare the competence to undergo embryogenesis of oocytes that grew in serum-free vs. FBS-containing medium. Approximately 70% of the oocytes underwent maturation regardless of whether the medium was serum-free or contained FBS. Of the mature ova grown in medium containing FBS, 53% cleaved to the two-cell stage after insemination compared with only 6% of the ova grown in serum-free medium. Addition of fetuin to the serum-free medium used for oocyte growth increased the frequency of cleavage to the two-cell stage. Of the embryos derived from oocytes that grew in FBS-containing medium, 70% completed the two-cell stage to blastocyst transition.(ABSTRACT TRUNCATED AT 250 WORDS)

摘要

本研究的首要目标是确定在无血清培养基中卵母细胞的生长是否会影响透明带对α-糜蛋白酶消化的溶解性,这是透明带“硬化”的一个指标,反映了精子穿透透明带的潜在能力。从12日龄小鼠的窦前卵泡中分离出卵母细胞-颗粒细胞复合体,并在含有5%胎牛血清(FBS)的培养基或无血清培养基中培养10天。在无血清培养基中生长的卵母细胞的透明带硬度是在体内生长的新鲜分离的生发泡(GV)期卵母细胞或在含FBS培养基中体外生长的卵母细胞的四倍。添加胎球蛋白可防止在无血清培养基中生长的卵母细胞透明带的硬化。第二个目标是比较在无血清培养基和含FBS培养基中生长的卵母细胞进行胚胎发生的能力。无论培养基是无血清的还是含有FBS,大约70%的卵母细胞都会成熟。在含FBS培养基中生长的成熟卵子中,53%在受精后分裂到二细胞期,而在无血清培养基中生长的卵子只有6%。在用于卵母细胞生长的无血清培养基中添加胎球蛋白可增加分裂到二细胞期的频率。在源自含FBS培养基中生长的卵母细胞的胚胎中,70%完成了从二细胞期到囊胚期的转变。(摘要截短至250字)

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