干细胞因子对骨髓间充质干细胞定向迁移的影响

Effect of stem cell factor on the directional migration of bone marrow-derived mesenchymal stem cells

目的初步探讨干细胞因子对大鼠骨髓间充质干细胞(bone marrow-derived mesenchymal stemcells,BMSCs)定向迁移的影响。方法密度梯度离心和贴壁培养法分离培养BMSCs,并予以传代。利用RT-PCR方法检测BMSCs对干细胞因子受体c-kit的表达,使用Transwell小室建立体外趋化迁移模型,评估干细胞因子对BMSCs定向迁移的影响以及p38丝裂原活化蛋白激酶通路(p38MAPK)抑制剂SB203580对干细胞因子诱导BMSCs迁移能力变化的影响。结果通过密度梯度离心和贴壁培养法获得了纯化的MSCs。RT-PCR证实BMSCs表达c-kit;干细胞因子可趋化体外模型中BMSCs通过聚碳酸酯膜向下室内迁移,在0～50ng/mL浓度范围内迁移细胞数量随干细胞因子的浓度增加而增加。加入SB203580后干细胞因子诱导的BMSCs迁移受到抑制。结论干细胞因子可以趋化BMSCs发生定向迁移,这一迁移过程的细胞内信号转导与p38MAPK通路有关。

[Objectives] To explore the effect of stem cell factor on the directional migration of rat bone marrowderived mesenchymal stem cells. [Methods] BMSCs were isolated, cultured, and passaged by density gradient eentrifugation and adherent culture method. The receptor of stem cell factor, c-kit, expressed on BMSCs was evaluated by RT-PCR method. Using Transwell inserts technique, we established in vitro model to study the impact of SCF on the migration of BMSCs and explore the regulatory role of SB203580, a p38MAPK inhibitor, on the SCF-indueed migrating capacity change of BMSCs. [Results] BMSCs were successive subculture and purified by density gradient centrifugation and adherent culture method. The results of RT-PCR showed that BMSCs expressed c-kit. SCF could induce BMSCs migrating towards lower compartment through polyearbonate membrane, and the number of migrating BMSCs increased with the increasing concentration of SCF from 0 ng/mL to 50 ng/mL. After adding SB203580, the migration of BMSCs was inhibited. [Conclusion] SCF can induce the migration of BMSCs, and p38MAPK signaling pathway is correlated with the intraeeilular signal transduction of this migration.