摘要
目的 :观察 5 氮杂胞苷 (5 Azacytidine)能否诱导骨髓间充质干细胞分化为心肌样细胞及其分化率 ,并探明骨髓间充质干细胞分化为心肌样细胞外 ,尚有哪些细胞类型存在 ?方法 :以成年SD大鼠为研究对象 ,分离单个核细胞 ,用 5 氮杂胞苷诱导骨髓间充质干细胞的分化。通过倒置显微镜观察受到诱导的骨髓间充质干细胞形态的变化 ,用免疫组化的方法标测分化的心肌样细胞的肌钙蛋白I(troponinI)和结蛋白 (desmin)是否存在 ?并用放免试剂盒测定分化的心肌样细胞能否分泌心房钠尿肽 ,透射电镜可以观察到分化的心肌样细胞内是否有肌丝和肌丝团的存在 ,RT PCR方法可以测定分化的心肌样细胞是否有Nkx2 .5和troponinI的cDNA的表达 ?从不同的层面鉴定诱导分化的心肌样细胞是否具有心肌细胞的特性和功能 ?并用流式细胞仪测定分化的心肌样细胞占细胞总数的百分比。结果 :① 5 氮杂胞苷可以诱导骨髓间充质干细胞分化为心肌样细胞。②在骨髓间充质干细胞分化为心肌样细胞的过程中 ,有部分成纤维细胞生长和未分化的单个核细胞存在。结论 :5 氮杂胞苷可以诱导骨髓间充质干细胞分化为心肌样细胞 ,5 氮杂胞苷诱导骨髓间充质干细胞分化为心肌样细胞后再做细胞移植 ,可能有改善受损心脏的功能。
AIM:Marrow stromal cells were isolated frow SD mice and were then induced by 5-azacytidine to confirm whether 5-azacytidine can induced marrow stromal cells into cardiomyocytes and the differentiation rate and to make sure of the cells involved in the course besides cardiomyocytes. METHODS: We use 2 month male SD mice were used as our subjects. Marrow stromal cells were obtoined by proportion 1.073 percoll. 24 hours later marrow stromal cells were treated with 5-azacytidine . Marrow stromal cells show round morphology and there is a single nucleus before treated. The morphology changed after the cells were treated with 5-azacytidine. They protruded pseudopods and connected with adjoining cells in one week and they connected together in 2～3 weeks. They grow clusterece forming muscle islands in four weeks. They secrectea atrial riatriuretic peptide. They were staina with anti-troponin-I and anti-desnim antinodies. Electron microscopy reveuled myotubes structure,and there were bridge-granules structures betreen adjoin cells. Tthese cells expressed Nkx2.5 and troponin I cDNA cardiomyocytes defferentiation was examined by flow cytometer . This cell line provided a powerful model for the study of cardiomyocytes differentiation. RESULTS: ① 5-azacytidine could induce bone marrow stromal cell into cardiomyocytes and the differentiation rate was about 25% . ②During marrow stromal cell differentiation, there was fibroblast cells and some round single nueclea cells. CONCLUSION : Our study shows that 5-azacytidine can induce bone marrow stromal cells into cardiomyocytes. After marrow stromal cells are treated with 5-azacytidine and grafted, they can improve damaged heart function.
出处
《心脏杂志》
CAS
2004年第6期505-508,512,共5页
Chinese Heart Journal
