小鼠精子冷冻的研究

Study on Cryopreservation of Mouse Spermatozoa

目的 探索、建立一套小鼠精子冷冻方法。方法 通过CPS和RS318两种冷冻液对比试验 ;进行不同解冻法试验 ;对B6 DF1 、CD 1、DBA、C57BL/ 6四种品系小鼠精子分别进行快速冷冻和缓速冷冻 ,将解冻精子、鲜精与卵母细胞进行体外受精 ,并进行体外培养、胚胎移植 ,通过受精率和受孕率对冷冻方法进行筛选。结果 两种冷冻液试验、不同解冻法试验的受精率差异均无显著性。B6 DF1 、CD 1、C57BL 6品系小鼠快速冷冻精子解冻后与卵母细胞的体外受精率 (35 %、34%、17% )显著低于鲜精 (6 0 %、5 9%、34% )、缓速冷冻精子的受精率 (6 2 %、5 8%、30 % ) ,此三种品系小鼠快速、缓速冷冻精子体外受精试验的受孕率均显著低于鲜精试验的受孕率。结论 缓速冷冻法是小鼠精子冷冻的适宜方法 ,干解冻法和稀释法处理冷冻液可简单有效地解冻精子。

Objective To explore and establish a suit of method of cryopreservation of mouse spermatozoa. Methods Two kinds of cryopreservation protection solutions(CPS and RS318) and two thawed methods were studied. Four strains(B 6DF 1,CD-1,DBA, C 57BL/6) of mouse spermatozoa were cooled rapidly and slowly respectively. Then frozen-thawed spermatozoa and oocytes were used for in vitro fertilization(IVF) under the control of fresh sperm. The techniques of culture and embryos transfer were used. Results There was no remarkable difference in the rates of IVF between CPS and RS318 or two thawed methods. The rates of IVF(35%,34%,17%)of tests of B 6DF 1,CD-I,C 57BL/6 mouse spermatozoa cooled rapidly were remarkably lower than those of fresh spermatozoa(60%,59%,34%) and spermatozoa cooled slowly(62%,58%,30%),and the rates of pregnancy in the tests of spermatozoa cooled were remarkably lower than those in fresh spermatozoa except for DBA. Conclusion Mouse spermatozoa cooled slowly is a feasible method, with dry thawed and dilution methods used.