不同来源骨髓间充质干细胞成骨能力的比较

目的 比较人不同来源骨髓间充质干细胞(human bone marrow mesenchymal stem cells,hBMSCs)的骨向分化能力,为骨组织工程筛选种子细胞提供一定的理论基础.方法 体外组织块培养法和有限稀释法培养颌骨来源骨髓间充质干细胞(jaw bone-marrow-derived mesenchymal stem cells,JMMSCs);骨髓穿刺法和密度梯度离心法培养长骨来源骨髓间充质干细胞(bone-marrow-derived mesenchymal stem cells,BMMSCs).流式细胞仪鉴定细胞表面抗原标记,成骨分化诱导后茜素红染色检测细胞矿化能力,成骨分化诱导后qRT-PCR及Western Blot检测细胞成骨相关分子:Runx2、1型胶原(Collagen Type 1,COL-1)、OCN.结果 JMMSCs和BMMSCs均阳性表达CD90、CD105,阴性表达CD34、CD14、CD45.成骨分化诱导21 d后,茜素红染色显示JMMSCs矿化能力强于BMMSCs.成骨诱导7 d、14 d、21 d后,JMMSCs成骨相关分子在基因和蛋白水平上均强于BMMSCs.结论 与BMMSCs相比,JMMSCs成骨分化能力较强,颌骨骨髓可能更适于用作骨组织工程的成体干细胞来源.

Comparison of osteogenic differentiation abilities of mesenchymal stem cells from different sources of hBMSCs

Objective To compare the osteogenic differentiation abilities of human bone marrow mesenchymal stem cells (hBMSCs) from different sources, and to provide basis for choosing a new source of seed cells in bone tissue engi-neering. Methods Jaw bone-marrow-derived mesenchymal stem cells (JMMSCs) were isolated from orthognathic surgi-cal sites and cultured by limited dilution for single cell clone. Long bone-marrow-derived mesenchymal stem cells (BMMSCs) were obtained from bone marrow of volunteers and isolated by density gradient centrifugation method. Flow cytometry was used to detect the surface markers of both cells. Osteogenic ability was assessed by PCR and Western Blot after osteogenic differentiation for the following molecules:Runx2, COL-1 and OCN. Alizarin red staining was used for determining the ability of cell mineralization after osteogenic differentiation. Results The expressions of cell sur-face markers CD90 and CD105 were positive in both type of cells, while CD34, CD14 and CD45 were all negative. After 21 days of osteogenic induction, JMMSCs formed significantly more mineralized nodules than BMMSCs. After 7, 14, 21 days of osteogenic induction, JMMSCs expressed more osteogenic-related molecules than BMMSCs. Conclusion The osteogenic differentiation capacity and mineralization ability of JMMSCs are significantly higher than BMMSCs. Jaw bone might be a more suitable source of seed cells in bone tissue engineering compared with long bone.