摘要
目的:建立骨髓间充质干细胞诱导培养成为破骨细胞的培养方法并进行初步的形态观察,为破骨细胞学及与其相关疾病的发病机制的研究提供更有意义的手段。方法:采用小鼠骨髓间充质干细胞通过1,25-(OH)2D诱导分化为破骨3细胞的培养方法,并观察原代成骨细胞对该方法破骨细胞形成及其功能的影响。根据是否加原代成骨细胞,分成未加成骨细胞、加成骨细胞二组,未加成骨细胞组是骨髓间充质干细胞直接诱导培养,加成骨细胞组是骨髓间充质干细胞与原代成骨细胞共同培养。分别培养6,9,12及15d,各组细胞到培养时间后取出进行TRAP染色阳性的多核细胞计数、骨片甲苯胺蓝染色及扫描电镜观察。结果:未加成骨细胞、加成骨细胞两组的骨髓间充质干细胞经培养后均出现破骨细胞,而且随着培养时间的延长,破骨细胞及骨片吸收陷窝的数目增多。培养相同时间的未加成骨细胞、加成骨细胞两组之间的破骨细胞及骨片吸收陷窝的数目差异无显著性意义。结论:通过1,25-(OH)2D诱导骨髓中的间充质干细胞培养破骨细胞的3方法是可行的,而且骨髓间充质干细胞诱导培养成破骨细胞时加入原代成骨细胞共同培养对破骨细胞的形成和骨吸收功能并无明显促进作用。
AIM:To investigate a method for culture of osteoclasts induced by bone marrow mesenchymal stem cells(MSCs) and observe the osteoclasts morphologically so as t o provide more significant ways for the study of osteoclasts and the pathogenesi s of osteoclast related diseases. METHODS:The culture method of osteoclasts induced and differentiated from mous e bone marrow MSCs by 1,25 dihydroxyvitamin D3[1,25 (OH)2D3]was used,and then the effect of primary osteoblasts on the formation and function of osteoclasts cultured with the above mentioned method was observed.According to whether prima ry osteoblasts were added,the culture was divided into two groups:bone marrow MS Cs were directly induced and cultured in non coculture group,and bone marrow MS Cs and primary cultured osteoblasts were cocultured in coculture group.On the 6t h,9th,12th and 15th day of culture,the number of osteoclasts was recognized as t artrate resistant acid phosphatase(TRAP)(+) multinucleate cells.The cellular TR AP(+) and the resorption lacuna on bone slice were examined with toluidine blue staining and under scanning electron microscope. RESULTS:Osteoclasts were found in the two groups after the culture of bone mar row MSCs.As the time prolonged,the number of osteoclasts and the number of resop tion lacuna on bone slice were increased,and no significant difference was found between the two groups at the same time of culture. CONCLUSION:Osteoclasts are cultured successfully from the bone marrow MSCs ind uced by 1,25 (OH)2D3.The formation of osteoclasts and bone resorption are not p romoted by the coculture of primarily cultured osteoblasts and bone marrow MSCs during culture of osteoclasts.
出处
《中国临床康复》
CSCD
北大核心
2005年第2期47-49,i002,共4页
Chinese Journal of Clinical Rehabilitation
