人骨髓基质干细胞定向诱导分化为成骨细胞及其体内成骨特征  

Induction and differentiation of human marrow mesenchymal stem cells into osteoblasts directionally and its features of osteogenesis in vivo

作　　者：殷晓雪[1] 陈仲强[1] 党耕町[1] 王志国[1] 张慧珠[1] 

机构地区：[1]北京大学第三医院骨科,北京市100083

出　　处：《中国临床康复》2005年第6期72-73,i004,共3页Chinese Journal of Clinical Rehabilitation

基　　金：国家自然科学基金资助(30271305)~~

摘　　要：目的:探讨应用人骨髓基质干细胞(humanmarrowmesenchymalseemcell,hMSC)进行骨科细胞治疗的可行性。方法:实验于2002-06/2003-02在北京大学第三医院骨科实验室完成。hMSC取自北京大学第三医院骨科脊柱外科手术患者,术前经检测无代谢性疾病。hMSC在诱导培养基的作用下定向分化为成骨细胞并大量扩增。RT-PCR法比较骨形成蛋白(BMP)-2mRNA在hMSC和成骨细胞中的表达差异。将成骨细胞复合PLGA植入裸鼠体内,硬组织切片,甲苯氨蓝染色,比较其与单纯植入成骨细胞或聚丙交酯-乙交酯(PLGA)时的异位成骨情况。结果:BMP-2mRNA在成骨细胞中的表达明显强于未经诱导分化的hMSC;成骨细胞与PLGA复合植入4周可见少量成骨,8周后可见成熟板层骨形成,PLGA组未见成骨,成骨细胞组8周仅可见少量成骨。结论:hMSC在诱导培养基的作用下可定向分化为成骨细胞并大量扩增。分化为成骨细胞后,BMP-2mRNA的表达显著升高。成骨细胞与PLGA复合植入裸鼠体内,能形成成熟的板层骨。hMSC在治疗骨缺损、骨不愈合等方面具有巨大的应用潜力。AIM:To study the feasibility of orthopedic cell therapy by means of human bone marrow mesenchymal stem cells(hMSCs). METHODS:The experiment was done in the orthopaedic laboratory,the Third Hospital of Peking University from June 2002 to February 2003.hMSC was harvested from donors undergoing spinal surgery in the Third Hospital of Peking University without any metabolic disease detected before operation.hMSC was differentiated into osteoblasts directionally and amplified greatly by culturing in an osteo inductive conditioned medium.The difference of bone morphogenic protein 2 (BMP 2) mRNA expression in hMSC and osteoblasts was compared by reverse transcriptase polymerase chain reaction(RT PCR).Osteoblasts were adhered to poly lactide co glycolide(PLGA) and then the compounds were implanted into the muscle of nude mice.The status of ectopia osteogenesis among compounds,PLGA and hOB were assayed by hard tissue section and toluidine blue staining. RESULTS:The expression of BMP 2 mRNA in osteoblasts was obviously stronger than that of uninduced and undifferentiated hMSC.Four weeks after implantation, a little bone like tissue was found in the group of compounds of osteoblasts and PLGA,while mature lamellar bone was observed after 8 weeks,but there was no bone tissue in the PLGA group, and only a little bone like tissue was found in the osteoblasts group after 8 weeks. CONCLUSION:hMSC can be induced and differentiated into osteoblasts directionally and greatly amplified by culturing in an osteo inductive conditioned medium.After hMSC were induced into osteoblasts, the expression of BMP 2 mRNA is significantly raised.Mature lamellar bone will form after the compounds of osteoblasts and PLGA are implanted into the muscle of nude mice.hMSC has great application potential in treating bone defect and nonhealing of bone.

关 键 词：骨髓 干细胞 骨生成 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]