不同年龄人体脂肪干细胞体外培养生长特点比较

Influence of age on the proliferation of adipose-derived stem cells in vitro

目的 观察不同年龄段人体脂肪干细胞（adipose-derived stem cells,ADSCs）体外培养的生长特点.方法 健康志愿者24例,年龄为26～45岁,按年龄分为4组：第1组6例,26～30岁;第2组6例,31～35岁;第3组6例,36～40岁;第4组6例,41～45岁.于不同年龄人体腹部皮下取用等量脂肪组织,胶原酶消化法分离、提取ADSCs,计数原代单核细胞数,记录细胞传代时间,观察传代细胞体外生长特点及检测细胞周期和细胞增殖活性.结果 4组不同年龄段原代单核细胞计数差异无统计学意义（P＞0.05）,而细胞生长至80％融合时所需时间,第2组短于第3组、第3组短于第4组（P＜0.05）;第3组细胞处于增殖期的比例明显高于第4组（P＜0.05）;前3组与第4组相比,四甲基偶氮噻唑蓝细胞活性较好,差异有统计学意义（P＜0.05）.结论 不同年龄段人体ADSCs体外培养细胞特性存在差异,随着年龄的增加细胞的增殖能力降低,其中＞40岁者的ADSCs增殖能力与年龄较小者相比,明显降低.

Objective In order to study the age associated changes,the characteristics of adipose derived stem cells （ADSCs） from 26 to 45 years old healthy volunteers were investigated.Methods Subcutaneous adipose tissue was obtained by tumescent liposuction from healthy women donors.ADSCs were isolated with collagenase digestion.We compared cells from 4 different age groups in 24 female patients ranged from 26 to 45 years old.Characteristics of growth and proliferation capacities of ADSCs were accessed in vitro.ADSCs were characterized by MTT and cell cycle detection methods.Results There was no significant difference of the mononuclear cells count between the four age groups while the first passage time of second and third group were shorter than that of third and fourth group,respectively （P＜0.05）.A significant increase in quiescence of S phase was observed in ADSCs from aged donors.ADSCs of third age group showed stronger capability than the fourth one （P＜0.05）.The ADSCs MTT activity of fourth group was significantly lower than the previous 3 groups （P＜0.05）.Conclusions Proliferation capacities of ADSCs in the group of older than 40 years decrease significantly compared with the younger groups.The results have significantly implicated that proliferation capacities of ADSCs are reduced with aging.