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目的 :建立人常染色体显性遗传型多囊肾病 (ADPKD)囊肿衬里上皮细胞系。 方法 :ADPKD囊肿衬里上皮细胞经FuGENE6介导转染pSV3质粒 ,并以G418筛选转染成功的细胞。转染后的第 48代细胞予以光镜、电镜形态学观察 ,细胞生长率测定 ,细胞染色体检查 ,碱性磷酸酶染色及细胞角蛋白、波形蛋白等免疫组化检查。结果 :第 48代细胞的碱性磷酸酶染色及抗细胞角蛋白、波形蛋白单克隆抗体反应阳性 ;形态学特征 ,细胞生长率与未经转化的第 2代细胞相比也无明显差异 ;但大部份转化细胞染色体为异倍体。 结论 :经鉴定 ,ADPKD囊肿衬里上皮细胞系已建立 ,可用于ADPKD细胞及分子生物学的研究。
Objective: To establish a cyst-lining epithelial cell line of autosomal dominant polycystic kidney disease (ADPKD). Methods: The pSV3 plasmids were transfected into the epithelial cells of ADPKD cyst lining by FuGENE6, and the transfected cells were screened by G418. The 48th generation cells were transfected with light microscopy, electron microscopy morphology, cell growth rate, cell chromosome examination, alkaline phosphatase staining and cytokeratin, vimentin and other immunohistochemical examination. Results: The 48th generation cells were positive for alkaline phosphatase staining and anti-cytokeratin and vimentin monoclonal antibody. The morphological characteristics and cell growth rate were also not significantly different from the non-transformed second generation cells Most of the transformed cells are aneuploid. Conclusion: It has been identified that ADPKD cyst-lining epithelial cell line has been established and can be used for ADPKD cell and molecular biology research.