中国畜牧兽医2017，Vol.44Issue(8)：2255-2260,6.DOI:10.16431/j.cnki.1671-7236.2017.08.006
驴皮成纤维细胞的体外培养研究
Study on in vitro Culture of Donkey Skin Fibroblasts
摘要
Abstract
This study was aimed to compare different culture methods for donkey skin fibroblasts to establish a highly efficient and rapid culture system.The dermal tissue of the healthy and 6-7 year old Xinjiang donkey was cultured for primary cells of skin fibroblast with tissue explants adherent method and double enzyme digestion method.Then,the growth characteristics,proliferation,cell cycle and mycoplasma contamination were detected and analyzed.The results showed that the fibroblast cells entered the logarithmic growth phase after cultured for 3 days by double enzyme digestion(0.25％trypsin digestion for 1 h and then 0.5％collagenase Ⅰ for 6 h),while it required 15 days with tissue explants adherent method.Flow cytometry analysis showed that nearly half of the cells were in G0/G1 stage,the other half in S+G2+M stage,while just 5.17％of the cells were apoptosis,which indicated that most of the cells were in the active stage of division and proliferation,and had strong vitality.Moreover,there was no mycoplasma contamination in the cultured cells.In summary,the culture system of donkey skin fibroblasts was rapidly and effectively established with the double enzyme digestion method in vitro.关键词
驴皮/成纤维细胞/组织块贴壁法/双酶消化法Key words
donkey skin/fibroblasts/tissue explants adherent method/double enzyme digestion method分类
生物科学引用本文复制引用
王艳萍,高帅,刘迥,曾维斌..驴皮成纤维细胞的体外培养研究[J].中国畜牧兽医,2017,44(8):2255-2260,6.基金项目
国家自然科学基金项目(31260531、31360530) （31260531、31360530）
兵团塔里木畜牧科技重点实验室项目(HS201503) （HS201503）
