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首页> 外文期刊>Experimental Animals >Development of Cyclodextrin-based Medium to Improve Fertilization Ability of Cryopreserved Sperm of C57BL/6 Mice
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Development of Cyclodextrin-based Medium to Improve Fertilization Ability of Cryopreserved Sperm of C57BL/6 Mice

机译：开发基于环糊精的培养基以提高C57BL / 6小鼠冷冻保存精子的受精能力

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摘要

Sperm cryopreservation provides an economical means of storing genetically engineered mouse strains in resource facilities. However, the fertilization rate for frozen/thawed sperm of C57BL/6, which is the main strain of genetically engineered mice, is very low. Therefore, it is necessary to establish an in vitro fertilization (IVF) method for cryopreserved C57BL/6 sperm that can obtain a high rate of fertilization after thawing. In the present study, we focused on the effects of methyl-β-cyclodextrin (M-β-CyD) on the fertilizing ability of frozen/thawed C57BL/6 sperm. Our results have shown that the highest fertilization rate for frozen/thawed sperm was obtained with M-β-CyD at 1.0 mM for 30 min (64%). The embryos with frozen/thawed sperm showed good developmental potential and the offspring had normal fertility. The efflux of cholesterol from frozen/thawed sperm was markedly increased by M-β-CyD in a dose-dependent manner. The effect of M-β-CyD occurred much earlier and to a greater extent than bovine serum albumin (BSA). The localization of cholesterol labeled by filipin in the sperm plasma membrane was drastically decreased by M-β-CyD. In summary, we suggest that M-β-CyD is useful for developing an IVF method for frozen/thawed C57BL/6 mouse sperm achieving a high fertilization rate, being involved in the capacity to sequester cholesterol from sperm membrane.
机译：精子冷冻保存提供了在资源设施中存储转基因小鼠品系的经济方法。然而，作为基因工程小鼠的主要品系的C57BL / 6的冷冻/解冻精子的受精率非常低。因此，有必要建立一种冷冻保存的C57BL / 6精子的体外受精（IVF）方法，该方法可在融化后获得较高的受精率。在本研究中，我们集中于甲基-β-环糊精（M-β-CyD）对冷冻/融化的C57BL / 6精子受精能力的影响。我们的结果表明，用1.0 mM的M-β-CyD30分钟（64％）可获得冷冻/解冻精子的最高受精率。具有冷冻/解冻精子的胚胎显示出良好的发育潜能，并且后代具有正常的受精能力。 M-β-CyD以剂量依赖性方式显着增加了冷冻/解冻精子中胆固醇的流出。与牛血清白蛋白（BSA）相比，M-β-CyD的作用发生得更早，程度更大。 M-β-CyD显着降低了由脂蛋白标记的胆固醇在精子质膜中的定位。总之，我们认为M-β-CyD可用于开发IVF方法，用于冷冻/解冻的C57BL / 6小鼠精子，实现高受精率，并涉及从精子膜中分离胆固醇的能力。

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