猪气管上皮细胞的分离、培养和鉴定

Growth and proliferous environment for porcine tracheal epithelial cell in vitro

目的探索猪气管上皮细胞体外适宜的生长、增殖条件,以期为组织工程化气管内壁上皮化提供种子细胞。方法无菌切取猪颈段气管,细菌蛋白酶消化。以明胶作为铺层,接种、培养。待细胞贴壁后,进行光镜观察、广谱角蛋白免疫组化检测、免疫荧光鉴定。结果光镜观察可见细胞呈铺路石样生长。角蛋白检测和免疫荧光检测均呈阳性,而阴性对照组和空白对照组呈阴性。结论以明胶作为铺层可以培养出活力较好、纯度高的气管上皮细胞。

Objective The purpose of this study was to explore the appreciate growth, pmliferons environment for porcine tracheal epithelial cell in vitro, which could be a new seed cell source for tissue engineered trachea epithelium. Methods Porcine tracheal epithelial cells were isolated by bacterial protease digestion and cultured on collagen foils Gelatin. The morphology of the cultured tracheal cells was evaluated by light microscopy and the keratin expression was analyzed by immunocytochemistry and immunofluorescence. Results The morphological observation confirmed the typical slabstoning shape of epithelial cell. Both kertin immtmocytochemistry and immtmofluorescence expression was positive, and the expression in blank control group was negative. Conclusion The culture system provide a ideal model for culturing tracheal epithelial cells, collagen foil Gelatin may play a important role in epithelial cell attachment.