摘要
目的:观察骨形成蛋白2(BMP-2)基因转染对骨髓间充质干细胞(BMSCs)碱性磷酸酶(ALP)活性和钙含量的影响。方法:成年大耳白兔16只,经髂后上嵴抽取骨髓组织混合后培养、分离BMSCs。取第3代细胞,采用电转法转染pcDNA3.1-BMP-2质粒。在扩大培养的第7、14、21、28和35天时收集细胞,用相应试剂盒对细胞ALP活性和钙含量进行测定,在扩大培养的第21天采用RT-PCR检测细胞BMP-2 mRNA的表达水平。以未转染细胞作对照。结果:转染组细胞BMP-2 mRNA相对表达水平为(1.652±0.082),对照组为(0.473±0.021),转染组BMSCs中BMP-2 mRNA的表达明显提高(t=2.192,P=0.005)。转染组细胞内ALP活性和钙含量较对照明显提高(P<0.01)。结论:转染BMP-2基因的BMSCs表现出明显的成骨活性,可以应用为骨组织工程研究的种子细胞。
Aim：To investigate the changes of alkaline phosphalase （ ALP） activity and calcium deposition of rabbit bone marrow mesenchymal stem cells （BMSCs） transfected with BMP-2 gene.Methods：The bone marrow were extracted from posterior superior iliac spine of 16 adult rabbits ,and mixed to be cultured .BMSCs were obtained and transfected with pcDNA3.1-BMP-2.At the 7th,14th,21st,28th and 35th day during the amplification culture ,ALP activity and calcium dep-osition were detected;at the 21st day, BMP-2 mRNA was detected using RT-PCR.The BMSCs without transfection were＆amp;nbsp;the control.Results：The expression of BMP-2 mRNA was （1.652 ＆#177;0.082） in transfected BMSCs, and （0.473 ＆#177;0.021） in the control BMSCs,and the difference between them was significant （t=2.192,P=0.005）.ALP activity and calcium deposition of transfected cells were higher than those of the control （P＆lt;0.01）.Conclusion：BMSCs transfected with BMP-2 gene show obvious osteogenic activity ,and could be used as seed cells for bone engineering research .
出处
《郑州大学学报（医学版）》
CAS
北大核心
2014年第1期102-105,共4页
Journal of Zhengzhou University(Medical Sciences)
