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目的 建立大鼠肺泡Ⅱ型上皮细胞 (简称AT Ⅱ )的改良体外原代培养法 ,并比较其鉴定方法的优劣。方法 AT Ⅱ分离、用胰蛋白酶消化法并用IgG包被的培养瓶粘附纯化 ,原代培养后通过鞣酸多彩、碱性磷酸酶 (Alkalinephosphatase ,AKP)、肺表面活性物质相关蛋白A(SP A)免疫组化三种染色法和电镜鉴定AT Ⅱ。结果 胰酶消化加IgG粘附纯化后所得的AT Ⅱ纯度为 90 % ,AKP染色鉴定最好。结论 采用IgG粘附纯化能提高AT Ⅱ纯度 ,AKP染色简便实用。
Objective To establish a modified primary culture method of rat alveolar type Ⅱ epithelial cells (AT Ⅱ), and to compare the advantages and disadvantages of the methods. Methods AT Ⅱ was isolated and purified by trypsin digestion and culture in IgG-coated flasks. After primary culture, the cells were stained with tannin, Alkaline Phosphatase (AKP), pulmonary surfactant associated protein A ) Immunohistochemistry three staining and electron microscopy AT Ⅱ. Results The purity of AT Ⅱ obtained after trypsin digestion and IgG adhesion purification was 90%, and AKP staining was the best. Conclusion Adherent IgG purification can improve the purity of AT Ⅱ, AKP staining is simple and practical.