C57-ras癌症转基因小鼠模型胚胎冷冻保种技术研究

目的 建立C57-ras癌症转基因小鼠模型胚胎的冷冻保种技术.方法 通过直接收集体内发育的2-细胞胚或者通过体外受精方法获得2-细胞胚,采用玻璃化冷冻法冷冻、复苏、移植,后代经PCR检测.结果 体内发育至2细胞期胚胎90枚,解冻后的平均复苏率是67.78％,移植产仔率是21.31％,后代的阳性率是23.08％；体外受精的2细胞期胚胎203枚,解冻复苏60枚,平均复苏率是63.33％,移植产仔率是15.78％,后代的阳性率是33.33％.结论 两种胚胎冷冻途径均获得了C57-ras癌症转基因动物模型阳性小鼠,建立了该模型动物的低温保存方法.

Study on the in vitro Fertilization and Embryo Cryopreservation in Carcinogenesis Transgenic C57-ras Mouse Model

Objective Investigate the application of in vitro fertilization （IVF） and embryo cryopreservation in C57-ras transgenie mice. Methods 2-cell embryos were collected in two ways, first of which is by 4-week superevulated C57BL/6J female mice mated with C57-ras transgentlc male mice, and the other is by IVF. The 2-cell embryos were cryopreserved after that. Then the cryopreserved 2-cell embryos were thawed and transplanted to caculate the ratio. Genetype of offsprings were analysed by PCR. Results The average recovery rate of cryopreserved normal 2-cell embryo was 67.78% and 21.31% were born after thawed 2-cell embryos transplanted. 23.08% of the offsprings were positive of human e-Ha-ras gene; the average recovery rate of eryopreserved IVF 2-cell was d3.33% , and 15.78% were born after transplanted, and 33.33% of the offspring were positive. Conclusion The results suggested that there was no significant difference in cryopreservation of normal and IVF C57-ras 2-cell embryos. The research provided the eryopreservation method for C57-ras transgenie mouse model.