冻融小鼠圆形精子细胞在体外受精中的应用研究

Application value of frozen-thawed mouse round spermatids in in vitro fertilization

目的:探讨冻融小鼠圆形精子细胞在体外受精中的应用价值。方法:体外培养获得的小鼠单倍体精子细胞冻融复苏后,经4',6-二眯基-2-苯基吲哚(DAPI)荧光染料标记后,行小鼠卵胞质内圆形精子细胞显微注射(ROSI)(冻融组,n=259),同时以体外培养获得的新鲜单倍体精子细胞行ROSI(新鲜组,n=238)作对照,比较两组精子细胞受精能力和后续胚胎发育能力。结果:体外培养获得的小鼠单倍体精子细胞冻融后存活率为(75.9±2.3)%。冻融组与新鲜组受精率(51.9%vs 55.7%)、2细胞率(51.0%vs 62.2%)、4~8细胞率(41.8%vs42.9%)及桑囊胚形成率(12.2%vs 21.4%)均无显著性差异(P>0.05)。结论:冻融复苏后的小鼠圆形精子细胞与冷冻前相似,具有一定的受精和胚胎发育能力。

Objective ： To investigate the application value of the frozen-thawed round spermatids of the mouse in in vitro fertili- zation （IVF）. Methods ： Haploid spermatids of the mouse obtained in vitro were divided into a frozen-thawed and a fresh group and oocytes were collected from 6 - 8 weeks old female mice. After diamidino-phenyl-indole （DAPI） staining, the oocytes were subjected to intracytoplasmic round spermatid injection （ROSI）, 259 in the frozen-thawed and 238 in the fresh group. Comparisons were made between the two groups in the capacities of fertilization and embryonic development. Results ： The survival rate of the frozen-thawed haploid spermatids was （75.9 ± 2.3） %. No statistically significant differences were observed between the frozen-thawed and fresh groups after ROSI in the rates of fertilization （51.9 vs 55.7%, P 〉 0.05 ）, 2-cell embryos （51.0 vs 62.2%, P 〉 0.05）, 4 -8-cell embryos （41.8 vs 42.9% , P 〉 0. 05 ）, or morula-blastocysts （ 12.2 vs 21.4%, P 〉 0. 05 ）. Conclusion ： Frozen-thawed round spermatids of the mouse are similar to fresh ones in their capacities of fertilization and embryonic development.