实验动物与比较医学2017，Vol.37Issue(2)：130-135,6.DOI:10.3969/j.issn.1674-5817.2017.02.009
树鼩角膜原代上皮细胞的分离培养、纯化与鉴定
Primary Isolation, Culture, Purification and Identification of Corneal Epithelial Cells in Tree Shrew
摘要
Abstract
Objectives To establish a stable technique for primary isolation,culture and purification and identification of tree shrew (Tupaia belangeri) corneal epithelial cells,and to provide a new experimental material for reserch of human ophthalmic corneal diseases.Methods The primary corneal epithelial cells from tree shrew were obtained by improved double digests method,the cells were purifed by method of cornea peeled off with transforming growth factor(TGF)-β inhibitors,achieve the purpose of identification by the method of immunofluorescence with keratin 3/12 antibody.Results The primary corneal epithelial cells from tree shrew with high activity and purity were obtained by double digests method.The corneal epithelial cells were purified food by TGF-β inhibitors and gradient digestion,and the purified cells was maintained a good epithelial cells shape in the subculture.The result of tree shrew corneal epithelial cells by immunofluorescence staining with keratin 3/12 monoclonal antibodies were shown positive.Conclusion An efficiency,simple and economic method was estabilshed for in vitro tree shrew corneal epithelial cells.The primary corneal epithelial cells from tree shrew with high activity and purity were obtained by double digests method,the subcultured cells was maintained a good epithelial cells shape.It will supply a new materials for eye diseases research.关键词
树鼩/角膜上皮细胞(CECs)/原代培养/优化/鉴定Key words
Tree shrew (Tupaia belangeri)/Corneal endothelial cells/In vitro/Primary culture/Optimization分类
生物科学引用本文复制引用
苗雨润,宋庆凯,匡德宣,陈玲霞,尹博文,李晓飞,代解杰..树鼩角膜原代上皮细胞的分离培养、纯化与鉴定[J].实验动物与比较医学,2017,37(2):130-135,6.基金项目
国家科技支撑计划项目(2014BAI01B00),云南省联合支持国家计划项目(2015GA009) （2014BAI01B00）
