BMP-2基因转染人成纤维细胞株KMB-17量效关系的研究

The Study of Dose-Effect Relationship on Human Fibroblasts Strain KMB-17 Trans Fected with Bone Morphogenetic Protein-2 Gene

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 -

Objective: To confirm the optimal transfection dose of plasmid DNA and liposome (Lipofectin) on BMP-2 gene transfect human fibroblasts strain KMB-17. Methods: The different amount of eukaryotic expression plasmid encoding for BMP-2 (pBK-B2) was transfected into KMB-17 cell by different amount of liposome. The transfection ratio was evaluated by immunohistochemistry staining, the effect of proliferation was measured by MTT colorimetric assay. These results were used to be confirmed the best transfection dose. Results: When the amount of Liopfectin was 5?μL and plasmid DNA was 2?μg, the highest expression ratio of BMP-2 gene was achieved, at the same time there isnt negative effect on cell proliferation. Conclusion: Using the optimal dose of plasmid DNA and liposome transfect BMP-2 gene, the most of KMB-17 cells can be transfected and there isnt negative effect on cells proliferation.