混合共培养条件下骨髓间充质干细胞与脂肪间充质干细胞成骨性能分析  

Osteogenic Differentiation Capacity of Mixed Cultured Bone Marrow Mesenchymal Stem Cells and Adipose-derived Mesenchymal Stem Cells

作　　者：王小伟 高春生 晏慧超 李勇光 

机构地区：[1]湖北省第三人民医院骨科,湖北武汉430033

出　　处：《实用骨科杂志》2017年第12期1091-1094,共4页Journal of Practical Orthopaedics

基　　金：湖北省科技计划项目(2013CKB030)

摘　　要：目的探讨混合共培养条件下骨髓间充质干细胞(bone mesenchymal stem cells,BMSC)和脂肪间充质干细胞(adipose-derived mesenchymal stem cells,ADSC)成骨分化潜能变化。方法无菌条件下获取BMSC和ADSC,纯化后倒置显微镜下观察细胞形态,对两种细胞分别进行成脂肪诱导、成软骨、成骨诱导,分别利用油红O染色、甲苯胺蓝染色、茜素红染色进行鉴定。实验分四组:A组:BMSC+ADSC未诱导(BMSC︰ADSC=1︰1);B组:BMSC诱导组;C组:ADSC诱导组;D组:BMSC+ADSC诱导组(BMSC︰ADSC=1︰1)。诱导组采用成骨诱导液进行成骨诱导,体外培养14d后Real-time PCR检测骨钙素(osteocalcin,OCN)及Runt相关转录因子2(Runx-2)基因表达情况,以及通过Western-blot法检测OCN及Runx-2蛋白表达情况。结果两种细胞在体外诱导后油红O染色、甲苯胺蓝染色、茜素红染色均为阳性,q-PCR结果显示D组OCN及Runx-2表达量明显高于A、B、C组(P<0.05),Westernblot结果显示OCN及Runx-2蛋白表达量D组明显高于其他三组P<0.05)。结论 BMSC与ADSC在体外混合共培养条件下可促进干细胞的成骨性能。Objective To assess the osteogenic differentiation in the condition of co-culture of BMSC and ADSC. Methods ADSC and BMSC obtained under sterile conditions. Oil red O was used to identify two cells after being in- duced into fat.Toluidine blue staining was used to identify two cells after being induced into Chondrocytes.Alizarin red staining were used in osteogenic induction group. Osteogenic induction group was divided into four sub-groups, group A：BMSC＋ADSC uninduced group（BMSC.. ADSC= 1 ： 1） ;group B： BMSC induced group; group C.. ADSC induced group ; group D： BMSC＋ ADSC induced group（BMSC： ADSC= 1 ： 1）. OCN and RUNX2 mRNA expression were de- tected by qPCR after osteogenic induction for 14 days,while the protein expression of OCN and RUNX2 were tested by Western blot at the same time.Results Oil red O staining, Toluidine blue staining and Alizarin red staining showed positive results.The mRNA expression and protein expression of OCN and RUNX2 were significiantly higher than the other three groups after osteogenie induction for 14 days in vitro. Conclusion The ability of osteogenic potential strengthened to some extent under the condition of co-culturing.

关 键 词：骨髓间充质干细胞 脂肪间充质干细胞 混合培养 成骨潜能 

分 类 号：R329.5[医药卫生—人体解剖和组织胚胎学]