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目的 :确定BMP - 2基因转染人成纤维细胞株KMB - 17的实验中质粒DNA和脂质体 (lipofectin)最佳使用剂量 ,以获得BMP - 2基因转染KMB - 17细胞的最佳条件 .方法 :采用不同剂量整合有BMP - 2基因的质粒DNApBK -B2和lipofectin转染KMB - 17细胞 ,通过免疫组化染色阳性细胞计数和MTT法检测细胞增殖的方法 ,确定不同剂量pBK -B2质粒DNA和lipofectin对细胞转染效率和增殖的影响 .结果 :当lipofectin用量为 5μL ,pBK -B2质粒DNA为 2 μg时 ,BMP - 2基因在KMB - 17细胞内表达比例最高 ,且对细胞增殖无明显影响 .结论 :采用合适剂量的质粒DNA和脂质体转染BMP - 2基因 ,可以使大多数KMB - 17细胞被转染并且不会影响细胞增殖
OBJECTIVE: To determine the optimal dosage of plasmid DNA and lipofectin in the experiment of transfecting human fibroblast cell line KMB - 17 with BMP - 2 gene in order to obtain the optimal conditions for BMP - 2 gene transfection into KMB - 17 cells Methods: KMB - 17 cells were transfected with plasmids pBK - B2 and lipofectin with different doses of BMP - 2 gene. The number of pBK - B2 plasmids was determined by immunohistochemical staining and MTT assay. DNA and lipofectin on the cell transfection efficiency and proliferation.Results: When the amount of lipofectin was 5μL and the pBK-B2 plasmid DNA was 2μg, the expression of BMP - 2 gene was the highest in KMB - 17 cells, Conclusion: Transfection of BMP - 2 gene with appropriate dose of plasmid DNA and liposome can make most of KMB - 17 cells transfected without affecting cell proliferation