饲养层种类及密度对体外培养兔原始生殖细胞的影响-中国细胞生物学.PDFVIP

中国细胞生物学学报 Chinese Journal of Cell Biology 2018, 40(8): 1384–1391 DOI: 10.11844/cjcb.2018.08.0044 技术与方法 饲养层种类及密度对体外培养兔原始生殖细胞的影响 王亚丹 虎啸 丁雪粉 朱邯豫 赵玉玺 吕海淼 彭展 王新庄* (河南农业大学牧医工程学院, 郑州 450002) 摘要 该研究以14~18天胎兔为试验对象, 体外分离培养扩增原始生殖细胞(primordial germ cells, PGCs), 观察其形态变化及集落形成过程。用AKP染色(alkaline phosphatase staining)及分子生 物学等方法鉴定兔PGCs, 结果均为阳性, 将兔PGCs分别接种于鼠胚胎成纤维细胞(mouse embryo 报 fibroblast, MEF) 、兔胚胎成纤维细胞(rabbit embryo fibroblast, REF)制成的不同密度饲养层来确定 种属跟密度对体外培养兔PGCs的影响。结果显示, 兔PGCs接种于用同源胚胎成纤维细胞所制成 4 的密度为6×10 的饲养层上所得到的集落个数最多, 集落形态最好, 分化速度较慢。 关键词 兔; 小鼠; 胚胎成纤维细胞; 饲养层; 原始生殖细胞 学 Effects of Feeder Layer Type and Density on Culturing 学 Rabbit (Leporidae) Primordial Germ Cells In Vitro Wang Yadan, Hu Xiao, Ding Xuefen, Zhu Hanyu, Zhao Yuxi, Lü Haimiao, Peng Zhan, Wang Xinzhuang* 物 (College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou 450002, China) Abstract The subject in the experiment was 14-18 d fetal rabbits. The primordial germ cells (PGCs) were 生 isolated and cultured in vitro. After observing the morphological changes of PGCs and colony forming process, we used the AKP staining (alkaline phosphatase staining) and molecular biological methods to identify rabbit PGCs. Inoculate PGCs to mouse, rabbit embryo fibroblast feeder layer made of different density to determine the effects of 胞 the species and density on the culture of rabbit primordial germ cells in vitro. The results showed that rabbit PGCs were inoculated on feeder layers with a density of 6×104 made from homologous embryonic fibr