建立稳定表达Smad蛋白的MDPC-23细胞模型

Generation of stable MDPC-23 cell lines overexpressing Smad proteins

建立稳定表达Smad蛋白的MDPC -2 3细胞克隆。方法 :将Smad表达载体转染进MDPC -2 3细胞内 ,通过G418筛选出阳性克隆 ,用Flag抗体进行Westernblot鉴定。 结果 :表达Flag融合蛋白细胞克隆为稳定整合Smad基因的阳性克隆 ,不表达Flag融合蛋白细胞克隆为表达空载体的细胞克隆。结论 :获得稳定表达Smad的细胞克隆 。

Objective:To establish MDPC-23 cell clones which stably overexpressed Smad proteins.Method:Transfection of Smad expression plasmids into MDPC-23 were done via the LipofectAMINE 2000 method. G418-resistant colonies were selected by adding G418 (300 μg/ml) to the medium for 2 weeks. Viable colonies were further screened by Western blot with anti-Flag M2 monoclonal antibody. Result: The cell clones which expressed the Flag protein are positive,while the cell clones which did not express the Flag protein are empty vector control.Conclusion:We successfully generate the MDPC-23 cell line which stably overexpressed Smad protein, which will help to investigate the role of Smad proteins in odontoblast differentiation.