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ADSCs3d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3d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36,48h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.
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 significantly higher than those in control group（ P 〈 0.05 or P 〈 0.01 ）. The apoptosis rate of HaCaT cells in experiment group was obviously 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 at 36 and 48 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.
出处
《第四军医大学学报》
北大核心
2009年第13期1206-1208,共3页
Journal of the Fourth Military Medical University
基金
国家自然科学基金(30772249)
关键词
脂肪干细胞
HACAT细胞
增殖
迁移
凋亡
adipose-derived stem cells
HaCaT cells
proliferation
migration
apoptosis
