基质细胞衍生因子-1对表皮干细胞的促增殖作用

Promoting Effect of Stromal Cell-derived Factor-1 on Proliferation of Epidermal Stem Cell

目的了解基质细胞衍生因子-1(SDF-1)在体外对表皮干细胞(ESCs)生长增殖的促进作用,为创面促愈药物研发提供理论依据。方法体外分离、培养ESCs,通过ESCs的免疫组化、免疫荧光染色对ESCs进行鉴定;RT-PCR检测ESCs中基质细胞衍生因子-1受体(CXCR4)基因的表达;取培养至第三代的ESCs分组,通过台盼蓝拒染实验、Edu标记法观察在不同剂量SDF-1的刺激作用和CXCR4拮抗剂存在下,A～E组及A’～E’组间ESCs生长增殖的差异。结果应用特制的ESCs培养基能培养出活力强、纯度高的ESCs;培养的ESCs表达K19、CD29、integrinα6,不表达CD71;CXCR4基因在ESCs中阳性表达;体外细胞实验证实SDF-1对ESCs有促进增殖作用,SDF-1浓度为100μg/L时作用最强,差异有统计学意义(P<0.01);CXCR4拮抗剂AMD3100存在时,SDF-1对ESCs的促增殖作用失效。结论 SDF-1对体外培养扩增的ESCs有促进增殖作用;SDF-1合理应用,有可能在皮肤治疗中发挥促愈作用。

Objective To explore the effect of stromal cell-derived factor-1（SDF-1） on proliferation of the epidermal stem cells（ESCs） in vitro,to provide an important theortical basis for development of new drugs which excels in promoting healing.Methods ESCs were isolated and cultured from healthy human foreskins which were identified by immunohistochemistry and immunofluorescence staining.The CXCR4 gene expression in ESCs was determined by RT-PCR.The passage cells were divided into several groups.The functions of the SDF-1 under different dose affected on ESCs in different groups was investigated by Trypan Blue staining and Edu incorporation experiment.Furthermore,the performances of ESCs in different groups were observed when they suffered from the CXCR4 antagonist.Results The ESCs were successfully cultured with special condition culture medium,and the cells expressed K19,CD29 and integrinα6,and negative in CD71.CXCR4 gene was also expressed in ESCs.The SDF-1 could promote the proliferation of the ESCs in vitro with dose dependent in a certain range,and the effect was most remarkable in 100μg/L groups（P0.01）.After adding the CXCR4 antagonist AMD3100 into the medium,SDF-1 no longer served any useful functions.Conclusion The SDF-1 may promote the proliferation of ESCs in vitro and it could be used in clinical treatment as a new drug for skin wound healing.