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在哺乳动物配子培养系统中用合成聚合物替代蛋白质。

Substitution of a synthetic polymer for protein in a mammalian gamete culture system.

作者信息

Bavister B D

出版信息

J Exp Zool. 1981 Jul;217(1):45-51. doi: 10.1002/jez.1402170106.

Abstract

Polyvinylalcohol (PVA) was tested as a replacement for protein (bovine serum albumin, BSA) in supporting motility, acrosome reactions, and fertilizing ability of hamster spermatozoa in vitro. Bovine serum albumin is normally required for all of these processes. After incubation for 5--6 hours in a simple culture medium containing BSA and PVA (0.1 mg/ml) and essential low molecular weight factors from blood serum, 85% of motile spermatozoa had undergone acrosome reactions. Sperm motility was equally well maintained by PVA in the absence of BSA but virtually no spermatozoa showed acrosome reactions even after prolonged incubation. Serum factors were later replaced by hypotaurine (10 microM), isoproterenol (1 microM), and penicillamine (20 microM). Spermatozoa incubated in this defined medium with BSA alone or with BSA and PVA fertilized more than 90% of oocytes. No oocytes were penetrated when BSA was replaced by PVA although vigorous sperm motility was maintained. Polyvinylalcohol may help elucidate the mechanism of the acrosome reaction by permitting effects of protein and other substances to be studied without loss of sperm motility (viability). Polyvinylalcohol could also replace BSA in solutions used for manipulation of zona pellucida-free oocytes. It is suggested that PVA may find general application in cell culture media.

摘要

聚乙烯醇(PVA)被用作蛋白质(牛血清白蛋白,BSA)的替代品,用于体外支持仓鼠精子的活力、顶体反应和受精能力。所有这些过程通常都需要牛血清白蛋白。在含有BSA和PVA(0.1毫克/毫升)以及血清中必需的低分子量因子的简单培养基中孵育5 - 6小时后,85%的活动精子发生了顶体反应。在没有BSA的情况下,PVA同样能很好地维持精子活力,但即使长时间孵育,几乎没有精子显示出顶体反应。血清因子后来被亚牛磺酸(10微摩尔)、异丙肾上腺素(1微摩尔)和青霉胺(20微摩尔)取代。在这种特定培养基中单独用BSA或用BSA和PVA孵育的精子使超过90%的卵母细胞受精。当用PVA取代BSA时,尽管精子活力旺盛,但没有卵母细胞被穿透。聚乙烯醇可以通过允许在不损失精子活力(生存能力)的情况下研究蛋白质和其他物质的作用,来帮助阐明顶体反应的机制。聚乙烯醇也可以在用于操作无透明带卵母细胞的溶液中取代BSA。有人认为PVA可能会在细胞培养基中得到广泛应用。

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