间充质干细胞条件培养液对人脐静脉内皮细胞增殖、迁移和黏附的影响

摘要

The aim of this study was to explore the effect of mesenchymal stem cell( MSC) conditioned medium (MSC-CM) on proliferation, migration and adhesion of human umbilical vein endothelial cell (CRL1730) and its mechanism. Isolation and purification of MSC were performed with the classic adhering method, the surface markers (CD29, CD90, CD45 and CD34) in MSC were detected by flow cytometry. MSC were treated and cultured for 3 d, the MSC-CM or MSC overexpressing stem cell-derived factor-1 (SDF-1) conditioned medium (Ad-SDF-1-MSC-CM) were collected. Subsequently, CRL1730 cells were treated respectively with 2% FBS-DMEM, 15% FBS-DMEM (control group), MSC-CM or Ad-SDF-1-MSC-CM for 24 h, the proliferation of CRL1730 cells was detected by MTT method. CRL1730 cell migration in vitro was performed by using wound healing system. The adhesion ability of CRL1730 cells was analyzed by microsecope . The results indicated that the CRL1730 cells treated with Ad-SDF-1-MSC-CM showed greater proliferative capacity than CRL173O cells treated with MSC-CM. While adding with AMD3100 5 ujnol/L, the blocker of CXCR4, the CRL1730 proliferation mediated by Ad-SDF-1-MSC-CM was significantly reduced. Meanwhile, compared with MSC-CM, Ad-SDF-1-MSC-CM had greater effects for promoting CRL1730 migration and enhancing adhesion ability of CRL1730 cells, these effects were significantly inhibited by AMD3100. It is concluded that MSC-CM promotes the migration and adhesion ability of CRL1730 cells through SDF-1 expressed by MSC.%本研究探索间充质干细胞(MSC)条件培养液对人脐静脉内皮细胞增殖、迁移和黏附能力的影响及其可能的机制.采用经典全骨髓贴壁法培养MSC,用流式细胞术鉴定MSC表面标志特征.收集MSC条件培养液(MSC-CM),过表达基质细胞细胞衍生因子1(SDF-1)的MSC条件培养液(Ad-SDF-1 -MSC-CM),以2％ FBS-DMEM、15％ FBS-DMEM培养液为对照,将MSC-CM、Ad-SDF-1 -MSC-CM等条件培养液加入人脐静脉内皮细胞株( CRL1730)作用24 h后,用MTT法检测CRL1730细胞的增殖情况,利用划痕法和黏附模型评价MSC-CM对CRL1730细胞迁移能力和黏附能力的影响.结果表明,与MSC-CM相比,Ad-SDF-1-MSC-CM处理的CRL1730细胞表现出更强的增殖能力,加入阻断剂AMD3100后,MSC-CM所介导的促增殖效应明显降低.MSC-CM、Ad-SDF-1-MSC-CM促进CRL1730细胞的迁移,AMD3100明显抑制其所介导的CRL1730细胞迁移.与MSC-CM相比,Ad-SDF-1 -MSC-CM处理的CRL1730细胞黏附能力明显增强,AMD3100能显著降低其增强CRL1730黏附能力的作用.结论:MSC-CM对CRL1730细胞的促增殖、迁移和黏附能力与其分泌的细胞因子SDF-1密切相关.