Development of genetic resources preservation for domestic animals using freeze-dried somatic cells

冷冻干燥体细胞可以克服目前冷冻保存方法的缺点，被认为是生产克隆动物的一种新工具。在此，我们研究了冷冻干燥对牛成纤维细胞DNA的损伤，以及FD细胞保存温度对移植后卵母细胞体外发育的影响。-30℃保存的FD细胞DNA损伤率为22%，+20℃保存的FD细胞DNA损伤率为100%。新鲜细胞、-30℃保存的FD细胞和+20℃保存的NT胚胎的卵裂率分别为67%、50%和0%。囊胚形成率分别为37%、28%和0%。保存在-30℃下的FD细胞与新鲜细胞在卵裂和发育到囊胚期方面没有显著差异。这些结果表明，从FD cell-NT卵母细胞中成功地产生了牛囊胚泡。

Freeze-dried somatic cells have been proposed as a new tool for production of clone animals, since it can overcome disadvantages of the current cryopreservation method. Here, we investigated the DNA damage on bovine fibroblast cells after lyophilization, and the influence of storage temperature of FD cells on in vitro development of oocytes after NT. The rate of DNA-damaged cells was 22% in FD cells preserved at -30C, and 100% at +20C. The cleavage rates of NT embryos with fresh cells, FD cells stored at -30C, and those stored at +20C were 67%, 50%, and 0%, respectively. The blastocyst formation rates were 37%, 28%, and 0%, respectively. No significant differences were found between FD cells stored at -30C and fresh cells with regard to cleavage and development to the blastocyst stage. These results suggest that bovine blastocysts are successfully produced from FD cell-NT oocytes.