犬皮肤成纤维细胞的分离、培养及鉴定

目的探索和建立适用于犬皮肤成纤维细胞的体外分离、培养及鉴定的技术方法。方法采用组织贴块培养法和胰蛋白酶、胶原酶Ⅰ联合消化法对犬皮肤成纤维细胞进行体外培养、传代。并对所培养的细胞进行倒置显微镜观察和苏木素-伊红染色,观察成纤维细胞形态,并对培养细胞行波形蛋白免疫荧光染色。结果倒置相差显微镜下可见长梭形细胞生长,苏木素-伊红染色可见细胞呈漩涡状、平行排列,第5代细胞免疫荧光检测波形蛋白(vimentin)表达阳性。结论建立了高效快速分离和稳定培养成纤维细胞的方法,为诱导犬心房纤维化提供了充足的种子细胞。

Isolation, Culture and Identification of Fibroblasts from Canine Skin

Objective To explore and set up the methodology of isolation,cultivation,and identification of skin fibroblasts in vitro.Methods Fibroblasts were released from dog skin by digestion of the dog skin with ype I collagenase and trypsin.The morphology and phenotype characterization of cultured fibroblasts was inspected by light microscopy after HE and immunocytochemical staining for vimentin.Results Fibroblasts exhibit spindle shape and arranged in swirl or parallel under the inverted phase-contrast microscope.Conclusions This study provides a technique of fast and efficient isolation and cultivation of skin fibroblasts,providing enough cell sources for dog atrial fibrosis research.