摘要
目的:研究骨形态形成蛋白9(Bone morphogenetic proteins9,BMP9)在C3H10T1/2干细胞向心肌细胞分化过程中的影响。方法:以高滴度(2.67ml/L)pAdEasy-BMP9重组腺病毒质粒转染C3H10T1/2干细胞,1周后应用RT-PCR方法及激光共聚焦方法分别检测C3H10T1/2干细胞中心肌特异转录因子及特异性蛋白的表达变化。2周后应用电子显微镜观察细胞超微结构的改变。结果:C3H10T1/2干细胞经pAdEasy-BMP9重组腺病毒质粒诱导1周后,细胞体积明显变大,排布走向趋于一致,细胞间连接紧密,折光性增强;细胞中开始出现心肌特异性转录因子NKx2.5、GATA-4、MEF2C及心肌特异性表达蛋白连接蛋白43(Connex-in43,Cx43)和心肌特异性肌钙蛋白T(Cardiac isoform of TropninT,cTnT)的表达;出现心肌特异性超微结构肌丝和闰盘。结论:BMP9可影响体外培养的C3H10T1/2干细胞定向分化为心肌样细胞。
Objective： To explore influence of BMP9 in myocardiocytes differentiation of stem cell In vitro. Methods：The C3H10T1/2 stem cell were transfected with 2.67ml/L pAdEasy-BMP9 Ad-GFP plasmid for 1 week and 2 weeks .The specific transcription factors of cardiomyocytes were detected in stem ceils during differentiation with plasmid transfection by RT-PCR and the specific proteins in cardiomy-ocytes were detected by laser confocal microsopy after 1 week ; the change of cells ultramicro structure were detected by electron microscope after 2 weeks. Results：Cells volume are increscent obviously,cells trend become unanimous,the connection between cells are compact, refractivity of cells enhance conspicuously after induction for 1 week.The expression of NKx 2.5, GATA-4, MEF2C could be detected in C3H10T1/2 stem cells with pAdEasy-BMP9 Ad-GFP plasmid tranfection. Cx43 and cTnT can be detected with plasmid tranfec- tion. Cardiomyocytes ultramicro structure can be detected after induction for 2 weeks. Conclusion：BMP9 makes a very important influence in C3H10T1/2 stem cells targeted differentiation into cardiomyocytes.
出处
《重庆医科大学学报》
CAS
CSCD
北大核心
2009年第6期672-676,共5页
Journal of Chongqing Medical University
基金
国家自然科学基金资助项目(30772361)
