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目的比较人不同来源骨髓间充质干细胞(human bone marrow mesenchymal stem cells,h BMSCs)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q RT-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。
OBJECTIVE: To compare the osteogenic differentiation of human bone marrow mesenchymal stem cells (hMSCs) from different sources, and to provide a theoretical basis for the screening of seed cells by bone tissue engineering. Methods The jaw bone-marrow-derived mesenchymal stem cells (JMMSCs) were cultured in vitro by tissue culture and limited dilution methods. Bone marrow-derived mesenchymal stem cells (JMMSCs) were cultured by bone marrow aspiration and density gradient centrifugation bone-marrow-derived mesenchymal stem cells, BMMSCs). Cell surface antigen labeling was identified by flow cytometry. Alizarin red staining was used to detect the mineralization ability of cells after induction of osteogenic differentiation. QRT-PCR and Western Blot were used to detect the osteogenesis-related molecules of Runx2 and collagen type 1 Collagen Type 1, COL-1), OCN. Results Both JMMSCs and BMMSCs had positive expression of CD90 and CD105, negative expression of CD34, CD14 and CD45. After 21 days of osteogenic differentiation induction, alizarin red staining showed that JMMSCs mineralized stronger than BMMSCs. After osteogenic induction for 7 d, 14 d, and 21 d, the osteogenesis-related molecules of JMMSCs were stronger than BMMSCs in gene and protein level. Conclusion Compared with BMMSCs, JMMSCs have strong ability of osteogenic differentiation. The bone marrow of the jawbone may be more suitable for the source of adult stem cells for bone tissue engineering.