CAJ | 학술논문

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。
목적：관찰골형성단백2（BMP-2）기인전염대골수간충질간세포（BMSCs）감성린산매（ALP）활성화개함량적영향。방법：성년대이백토16지，경가후상척추취골수조직혼합후배양、분리BMSCs。취제3대세포，채용전전법전염pcDNA3．1-BMP-2질립。재확대배양적제7、14、21、28화35천시수집세포，용상응시제합대세포ALP활성화개함량진행측정，재확대배양적제21천채용RT-PCR검측세포BMP-2 mRNA적표체수평。이미전염세포작대조。결과：전염조세포BMP-2 mRNA상대표체수평위（1．652±0．082），대조조위（0．473±0．021），전염조BMSCs중BMP-2 mRNA적표체명현제고（t＝2．192，P＝0．005）。전염조세포내ALP활성화개함량교대조명현제고（P＜0．01）。결론：전염BMP-2기인적BMSCs표현출명현적성골활성，가이응용위골조직공정연구적충자세포。
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 the control.Results:The expression of BMP-2 mRNA was (1.652 ±0.082) in transfected BMSCs, and (0.473 ±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<0.01).Conclusion:BMSCs transfected with BMP-2 gene show obvious osteogenic activity ,and could be used as seed cells for bone engineering research .