摘要
研究了牛胎儿组织成纤维细胞的分离、培养、纯化方法和生长特征,并对培养细胞的冷冻保存和复苏进行了观察。采取组织块培养法进行原代培养,在接种第5天到第6天成纤维细胞生长旺盛;用0.25%胰蛋白酶消化,传代培养细胞生长状态良好;用液氮冷冻法保存传代细胞,解冻后持续传代至第12代仍生长良好,第8代细胞冻存前和复苏后的活率分别为97.0%和94.3%,无显著差异(P>0.05);分离纯化的胎牛成纤维细胞的生长曲线都正常;成功地建立了牛胎儿成纤维细胞系。该细胞可经多次传代培养和冷冻保存。
The bovine fetal fibroblasts isolation and culture,purification methods and growth characteristics were studied,and the cryopreservation of cultured cells and recovery were observed.Bovine fetal fibroblasts were isolated by attaching tissue explants from ear skin of a bovine fetal,inoculated with 5～6 days fibroblast growth vigorous.Fibroblasts with 0.25% trypsin digestion and subcultured cells in good condition.With liquid nitrogen freezing method to save passage fibroblasts,fibroblasts were thawed into a continuous passage to post～12 generation continues to grow well.8 passages fibroblasts before and after cryopreservation of live after resuscitation rates were 97.0% and 94.3%,no significant difference （P= 0.06）.Separation and purification of fetal bovine fibroblasts growth curves were normal.The bovine fetal fibroblasts line was successfully established.The bovine fetal fibroblasts can be subcultured several times and cryopreservation.
出处
《中国草食动物》
2010年第3期5-8,共4页
China Herbivores
关键词
牛胎儿
成纤维细胞
分离培养
冷冻保存
bovine fetal
fibroblast cell
isolation and culture
cryopreservation
