藏野驴皮肤成纤维细胞分离培养及生物学特性分析

Isolation,Culture and Biological Characteristics of the Fibroblasts from the Skin Tissue of Equus kiang

为了研究藏野驴成纤维细胞分离培养方法及生物学特性,以7岁龄藏野驴耳部皮肤组织为试验材料,采用组织块贴壁法进行细胞原代培养,采用胰酶消化法和差速贴壁法进行细胞纯化,通过细胞形态观察、生长曲线测定、细胞周期和免疫荧光染色对细胞进行鉴定。结果显示,成纤维细胞生长曲线呈“S”形,细胞生长状态良好,细胞特异性标记免疫荧光染色显示FSP-1呈阳性表达。结果表明,成功建立了藏野驴成体耳部皮肤成纤维细胞体外分离、培养和鉴定方法,为藏野驴种质资源保护、细胞水平及分子水平的研究奠定了基础。

In order to study the isolation,culture methods and biological characteristics of the fibroblasts of Equus kiang,the ear skin tissue of a 7-year-old Equus kiang was used.The primary cells were obtained by adherent method.The cells were purified with trypsin digestion method and a technique of differential anchoring velocity.The cells were identified by cellular morphology,growth curve,cell cycle and immunofluorescence staining.The results showed that the growth curve of fibroblasts was S-shaped,and the cells grew well.The expression of FSP-1 was confirmed positive by immunofluorescence staining.This study established the method of isolation,culture and identification of skin fibroblasts from ears skin of adult Equus kiang in vitro,which laid a foundation for the protection of germplasm resources in Equus kiang and the research at the cellular and molecular levels.