摘要
目的 研究纯化脐血CD34+细胞移行穿越血管内皮细胞能力及影响因素。方法 免疫微磁珠阳性选择法(MACs)纯化脐血CD34+细胞,流式细胞仪测定基质细胞衍生因子(SDF-1α)受体(CXCR4)表达率;与干细胞因子(SCF)、白介素(IL)-6,IL-3及flt3配体(FL)共同孵育12小时,静脉内皮细胞株(ECV)接种于transwell滤膜上层,研究CD34+细胞在SDF-1α作用下移行穿越ECV的能力,transwell滤膜孔径为8μm,并观察阻断其表面粘附分子(CD62L、CD62E和VLA-4)后对迁移的影响。结果 CD34+细胞在自然状态下也能少量通过ECV细胞层,SDF-1α可显著提高CD34+细胞移行能力,通过率与CD34+细胞表面CXCR4表达相关;CD34+细胞与SCF及IL-6共同孵育后,可明显提高其穿越ECV细胞的能力(P<0.01);单独或联合应用抗粘附分子抗体显著减少CD34+细胞的穿内皮细胞移行(P<0.01)。结论 CD34+细胞可穿过ECV内皮细胞层向SDF-1α浓度高的一侧移行,与IL-6和SCF共同培养后可增强CD34+细胞的移行能力,抗粘附分子单抗显著减少CD34+细胞的移行。
Objective To investigate the transendothelial migration of cord blood CD34+ cells and effects of adhesion moleculer and hemalopoietic growth factor on the process. Methods MACs were employed to isolate the CD34+ cells from cord blood and the expression of CXCR4 on CD34+ was determined by flow cytometry. The purified CD34+ cells were cultured with stem cell factor (SCF) , interleukin(IL)-6,IL-3 and flt3 ligand(FL) for 12 hours. Endothelial cell line(ECV)was seeded on the upper compartment of trans well (pore size 8. 0μm). The number of migration was counted at different time section when the cells were incubatad with or without anti cell adhesion molecule (CAM) monoclone antibody. Results A small portion of CD34+ cells migrated through the ECV cell line spontaneously;SDF-1α significantly increased transendothelial migration (P<0. 01). The percentage of migration correlated with the expression of CXCR4 on CD34+ cells. Transendothelial migration greatly enhanced when CD34+ cells were incubated with SCF and IL-6 (P<0. 01). Conclusion CD34+ cell migration through the ECV lines can be induced by SDF-1α. Incubation with IL-6 and SCF promotes the transendothelial migration due to increased expression of CXCR4 on CD34+ cells,other wise, the migration was blocked with anti CAM antibodies.
出处
《江苏医药》
CAS
CSCD
北大核心
2003年第4期266-268,共3页
Jiangsu Medical Journal
基金
江苏省135重点医学人才基金(RC2002021)
苏州大学医学发展基金(EE123002)
