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目的观察体外长期培养对人脐血间充质干细胞(Human umbilical cord blood derived mesenchymal stem cells,UCB-MSCs)成骨分化潜能的影响,探讨其作为组织工程骨种子细胞的可行性。方法流式细胞技术观察长期体外培养的UCB-MSCs细胞表面抗原表达的变化规律,并通过Alizarin Red染色及碱性磷酸酶活性、骨钙蛋白含量和钙离子含量的检测观察长期体外培养对UCB-MSCs成骨特性的影响。结果第10代之前的UCB-MSCs能够高表达间充质干细胞表面标志物,7代之前的细胞体外增殖能力不随传代次数而发生明显变化。Alizarin Red染色及碱性磷酸酶活性、骨钙蛋白含量和钙离子含量的检测显示8代之前的UCB-MSCs仍能保持较强的成骨分化能力。结论7代之前的UCB-MSCs能保持稳定的体外成骨分化特性,有望成为较理想的组织工程骨种子细胞。
Objective To observe the effect of long-term culture in vitro on the osteogenic potential of human umbilical cord blood derived mesenchymal stem cells (UCB-MSCs) and to explore its feasibility as a seed cell for tissue engineering bone. Methods Flow cytometry was used to observe the changes of surface antigen expression on long-term cultured UCB-MSCs. Alizarin Red staining and alkaline phosphatase activity, osteocalcin content and calcium ion content were used to observe the effect of long-term culture on UCB- Effect of MSCs on osteogenic properties. Results UCB-MSCs before passage 10 were able to express mesenchymal stem cell surface markers with high levels. The proliferation ability of UCB-MSCs before passage 7 did not change significantly with the passage number. Alizarin Red staining and alkaline phosphatase activity, osteocalcin content and calcium content of the test showed that 8 generations before UCB-MSCs still maintain a strong osteogenic differentiation. Conclusion UCB-MSCs before 7 generations can maintain stable in vitro osteogenic differentiation characteristics, which is expected to become a more ideal tissue engineered bone seed cells.