Primary Isolation, Culture, Purification and Identification of Corneal Epithelial Cells in Tree Shrew

doi:10.3969/j.issn.1674-5817.2017.02.009

Abstract

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 .

Key words: Tree shrew (Tupaia belangeri), Corneal endothelial cells, In vitro, Primary culture

CLC Number:

Q95-33

MIAO Yu-run, SONG Qing-kai, KUANG De-xuan, CHEN Ling-xia, YIN Bo-wen, LI Xiao-fei, DAI Jie-jie. Primary Isolation, Culture, Purification and Identification of Corneal Epithelial Cells in Tree Shrew[J]. Laboratory Animal and Comparative Medicine, 2017, 37(2): 130-135.

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