人脂肪干细胞分泌因子对HaCaT细胞的生物学作用
Effects of conditioned medium of human ADSCs on HaCaT cells
折涛 1刘佳琦 1张军 1张万福 1蔡维霞 1白哓智 1胡大海1
作者信息
- 1. 第四军医大学西京医院全军烧伤中心,陕西,西安,710033
- 折叠
摘要
目的:分离培养脂肪干细胞(ADSCs)以探讨其分泌因子对HaCaT细胞的生物学作用.方法:从人腹部脂肪组织中分离并培养ADSCs 3 d后,ELISA法测定ADSCs分泌血管内皮生长因子(VEGF),肝细胞生长因子(HGF)含量;收集ADSCs培养液(ADSC-CM)作用于HaCaT细胞株.采用MTT法测定HaCaT细胞的增殖;AnexinV/PI双染色法测定HaCaT细胞的凋亡情况;体外细胞划痕法测定其迁移能力.结果:培养获得ADSCs细胞3 d后,培养液中VEGF、HGF含量分别为(287±47),(577±85)ng/L.MTT实验30%ADSC-CM组、50%ADSC-CM组的A490nm值分别高于对照组,差异具有统计学意义(P<0.05或P<0.01).实验组HaCaT细胞凋亡率明显低于对照组(P<0.05).与对照组相比较,实验组HaCaT细胞迁移距离36,48 h时间点显著增大(P<0.05或P<0.01).结论:ADSCs能分泌细胞因子且对HaCaT细胞具有促增殖、迁移和抑制凋亡作用.
Abstract
AIM: To isolate and culture adipose-derived stem cells(ADSCs) and to study the effects of cytokines secreted by ADSCs on HaCaT cells. METHODS: ADSCs were isolated from human adipose tissues and cultured for 3 d. ELISA was used to detect the secretion level of VEGF and HGF. The conditioned medium of ADSCs (ADSC-CM) was then used to culture HaCaT cells. MTT method was used to determine the proliferation and Annexin V/PI was used to examine the apoptosis rate of HaCaT cells. The cell migration was measured by in vitro wound-healing assay. RESULTS: ADSCs were isolated and cultured successfully. The amount of VEGF and HGF was (287±47) ng/L and (577±85) ng/L, respectively. The A490nm values of HaCaT cells in 30% ADSC-CM group and 50% ADSC-CM group were signifi-cantly higher than those in control group (P<0.05 or P<0.01). The apoptosis rate of HaCaT cells in experiment group was obvi-ously lower than that in control group (P<0.05). The migration distance of HaCaT cells in experiment was significantly longer than that in control group at36 and48 h(P<0.05 or P<0.01). CONCLUSION: ADSCs can secrete cytokines and ADSC-CM promotes proliferation and migration and suppresses apoptosis of HaCaT cells.
关键词
脂肪千细胞/HaCaT细胞/增殖/迁移/凋亡Key words
adipose-derived stem cells/HaCaT cells/prolifera-tion/migration/apoptosis引用本文复制引用
出版年
2009
NETL
NSTL
维普
万方数据