目的 通过诱导人白色前脂肪细胞(human white adipose tissue stromal vascular fraction,hWAT-SVF)分化为成熟脂肪细胞,检测不同分化周期中hWAT-SVF细胞成脂标志物的表达水平,为完善和优化人前脂肪细胞分化模型提供数据支持.方法 将hWAT-SVF细胞培养6天至融合状态,采用3-异丁基-1-甲基黄嘌呤、地塞米松、胰岛素等诱导细胞分化,在分化的第0、3、6、9、12天收集细胞,观察细胞大小和形态等变化,检测脂滴、中性脂质和甘油三酯(triglyceride,TG)的生成情况,以及脂肪生成相关基因和蛋白[过氧化物酶体增殖物激活受体γ(peroxisome proliferator-activated receptor γ,PPARγ)、脂肪酸结合蛋白4(fatty acid binding protein 4,FABP4)和脂滴包被蛋白1(perilipin 1,PLIN1)]等的表达水平.结果 经诱导分化后,hWAT-SVF细胞形态明显变大变圆,脂滴和TG含量显著升高,成脂标志物如PPARγ、FABP4、PLIN1等的表达水平显著上调,表明成功将hWAT-SVF细胞诱导为成熟脂肪细胞.分化第3天PPARγ;以及分化第9天成熟脂肪细胞标志物(FABP4和PLIN1)蛋白表达水平分别显著升高1.8、21、14.3倍.结论 在分化第9天,hWAT-SVF细胞已具备成熟脂肪细胞的特征.本研究成功缩短了人源前脂肪细胞分化模型的构建周期,为研究脂肪生成效应及机制提供了一种有效的细胞模型.
Effect of different induction stages on the differentiation level of human white preadipocytes
Objective Inducing human white adipose tissue stromal vascular fraction(hWAT-SVF)into mature adipocytes,to measure the expression levels of adipogenic markers during different stages of differentiation,and to provide support data for the im-provement and optimization of human preadipocyte differentiation models.Methods hWAT-SVF cells were cultured for 6 days until confluence.Cell differentiation was induced by 3-isobutyl-1-methylxanthine,dexamethasone,insulin,etc.Cells were collected on days 0,3,6,9,and 12 of differentiation.Changes in cell size and morphology were observed.The formation of lipid droplets,neutral lip-ids,and triglyceride was determined.The expression levels of adipogenic genes and proteins[e.g.,peroxisome proliferator-activated receptor γ(PPARy),fatty acid binding protein 4(FABP4),and perilipin 1(PLIN1)]were measured.Results After differentia-tion,the size of hWAT-SVF cells increased and their shape was closer to round,the contents of lipid droplets and triglyceride were sig-nificantly increased,and the expression levels of adipogenic markers PPARγ,FABP4,and PLIN1 were significantly upregulated.These results indicated that hWAT-SVF cells were successfully induced into mature adipocytes.The expression of PPARγ on day 3 of differentiation and the expression of FABP4 and PLIN1 as mature adipocyte markers on day 9 were significantly increased by 1.8,21,and 14.3 times,respectively.Conclusion On day 9 of differentiation,hWAT-SVF cells exhibited the characteristics of mature adipo-cytes.This study successfully shortened the construction period of human preadipocyte differentiation model and provided an effective cell model for studying the effect and mechanism of adipogenesis.
white adipocyteadipocyte differentiationadipogenic marker
万方数据
维普
