Yokoyama M, Akiba H, Katsuki M, Nomura T
Central Institute for Experimental Animals, Kawasaki-shi, Japan.
Jikken Dobutsu. 1990 Jan;39(1):125-8. doi: 10.1538/expanim1978.39.1_125.
Spermatozoa from cauda epididymis of mature mice were suspended in preservation solution (Dulbecco's PBS containing raffinose in combination with glycerol, DMSO or skim milk as freezing protective agents). The suspension was frozen by the dry ice-alcohol method and preserved for 1-120 days in liquid nitrogen (-196 degrees C). Highest sperm viability after thawing was obtained with a combination of 10% raffinose and 5% glycerol or with a combination of 10% raffinose and 10% DMSO. These frozen thawed sperm were found to have fertilizing capacity when used for in vitro fertilization. The 2-cell embryos obtained through the above procedures developed into normal pups at a high rate when transferred into the oviducts of pseudopregnant female mice.
将来自成熟小鼠附睾尾部的精子悬浮于保存液中（含有棉子糖并结合甘油、二甲基亚砜或脱脂乳作为冷冻保护剂的杜氏磷酸盐缓冲液）。通过干冰-酒精法对悬浮液进行冷冻，并在液氮（-196℃）中保存1至120天。解冻后精子活力最高的情况是10%棉子糖与5%甘油的组合，或10%棉子糖与10%二甲基亚砜的组合。发现这些冷冻解冻后的精子用于体外受精时具有受精能力。通过上述程序获得的2细胞胚胎转移到假孕雌性小鼠的输卵管中时，能以高比例发育成正常幼崽。
