小鼠精子的长期冷冻保存
Long-term cryopreservation of mouse sperm.
作者信息
Kaneko Takehito, Yamamura Ayako, Ide Yukie, Ogi Mami, Yanagita Tomoko, Nakagata Naomi
机构信息
Division of Reproductive Engineering, Center for Animal Resources and Development (CARD), Kumamoto University, 2-2-1 Honjo, Kumamoto 860-0811, Japan.
出版信息
Theriogenology. 2006 Sep 15;66(5):1098-101. doi: 10.1016/j.theriogenology.2006.02.049. Epub 2006 Apr 18.
The objective was to determine if mouse sperm can maintain their fertilizing ability after being frozen for >10 y and whether the offspring derived from these sperm had normal fertilizing ability and phenotype. We cryopreserved sperm from six strains of mice (C57BL/6J, DBA/2N, BALB/cA, C3H/HeJ, B6D2F1 and B6C3F1) in a solution containing 18% (w/v) raffinose and 3% (w/v) skim milk, and preserved them in liquid nitrogen for >10 y. To assess the normality and fertilizing ability of these sperms, they were thawed and used for in vitro fertilization of oocytes of the same strains. Fertilization rates for C57BL/6J, DBA/2N, BALB/cA, C3H/HeJ, B6D2F1 and B6C3F1 were 66.4, 92.3, 72.8, 32.9, 60.3 and 53.7%, respectively. Furthermore, 38.3, 15.0, 43.3, 26.1, 38.3 and 16.7% of the embryos transferred to pseudopregnant females developed and produced live offspring that had normal phenotype and fertility.
目的是确定小鼠精子在冷冻超过10年后是否仍能保持其受精能力，以及源自这些精子的后代是否具有正常的受精能力和表型。我们将来自六个品系小鼠（C57BL/6J、DBA/2N、BALB/cA、C3H/HeJ、B6D2F1和B6C3F1）的精子保存在含有18%（w/v）棉子糖和3%（w/v）脱脂乳的溶液中，并在液氮中保存超过10年。为了评估这些精子的正常性和受精能力，将它们解冻并用于对同品系的卵母细胞进行体外受精。C57BL/6J、DBA/2N、BALB/cA、C3H/HeJ、B6D2F1和B6C3F1的受精率分别为66.4%、92.3%、72.8%、32.9%、60.3%和53.7%。此外，移植到假孕雌性体内的胚胎中有38.3%、15.0%、43.3%、26.1%、38.3%和16.7%发育并产下了具有正常表型和生育能力的活后代。
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