通过去除培养基中的Ca2+显著提高冷冻保存的C57BL/6J小鼠精子的受精能力。

Marked improvement of fertility of cryopreserved C57BL/6J mouse sperm by depletion of Ca2+ in medium.

作者信息

Suzuki-Migishima Rika, Hino Toshiaki, Takabe Miho, Oda Kanako, Migishima Fujio, Morimoto Yoshiharu, Yokoyama Minesuke

机构信息

Mitsubishi Kagaku Institute of Life Sciences, Tokyo, Japan.

出版信息

J Reprod Dev. 2009 Aug;55(4):386-92. doi: 10.1262/jrd.20163. Epub 2009 Apr 13.

DOI:10.1262/jrd.20163

PMID:19367085

Abstract

Cryopreservation of mouse sperm is useful for maintaining various strains. However, fertility generally decreases after freezing. In particular, the fertility of cryopreserved C57BL/6J sperm is very low. To improve the fertility of frozen sperm, we examined the efficiencies of various media used for sperm preincubation (SP) and in vitro fertilization (IVF) in frozen C57BL/6J sperm. In this study, SP medium was examined for efficiency of fertility with respect to content, especially calcium (Ca(2+)), phosphate (PO(4)(3-)) and lactate. In all media containing no Ca(2+), including medium lacking Ca(2+), lacking Ca(2+) and PO(4)(3-), lacking Ca(2+) and lactate and lacking Ca(2+), PO(4)(3-) and lactate, high IVF rates were obtained (79, 69, 76 and 71%, respectively). On the other hand, the rates for media containing Ca(2+) were significantly lower (30-38%, P<0.05). After transfer, 41-50% of newborns were obtained in all media containing no Ca(2+). In conclusion, preincubation of thawed sperm in medium containing no Ca(2+) markedly improved the fertility of cryopreserved C57BL/6J sperm. These results indicate that the present method of IVF using medium with no Ca(2+) is practical for use in cryopreserved C57BL/6J sperm.

摘要

小鼠精子的冷冻保存对于维持各种品系很有用。然而，冷冻后生育能力通常会下降。特别是，冷冻保存的C57BL/6J精子的生育能力非常低。为了提高冷冻精子的生育能力，我们研究了用于冷冻的C57BL/6J精子预孵育（SP）和体外受精（IVF）的各种培养基的效率。在本研究中，针对培养基的成分，特别是钙（Ca(2+)）、磷酸盐（PO(4)(3-)）和乳酸，检测了SP培养基的生育效率。在所有不含Ca(2+)的培养基中，包括缺乏Ca(2+)的培养基、缺乏Ca(2+)和PO(4)(3-)的培养基、缺乏Ca(2+)和乳酸的培养基以及缺乏Ca(2+)、PO(4)(3-)和乳酸的培养基，均获得了较高的IVF率（分别为79%、69%、76%和71%）。另一方面，含Ca(2+)的培养基的IVF率显著较低（30 - 38%，P<0.05）。移植后，在所有不含Ca(2+)的培养基中均获得了41 - 50%的新生小鼠。总之，将解冻后的精子在不含Ca(2+)的培养基中预孵育可显著提高冷冻保存的C57BL/6J精子的生育能力。这些结果表明，目前使用不含Ca(2+)培养基的IVF方法对于冷冻保存的C57BL/6J精子是实用的。