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 improvement 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 lipids, and triglyceride was determined. The expression levels of adipogenic genes and proteinse.g., peroxisome proliferator-activated receptor γ (PPARγ), fatty acid binding protein 4 (FABP4), and perilipin 1 (PLIN1) were measured.

Results After differentiation, the size of hWAT-SVF cells increased and their shape was closer to round, the contents of lipid droplets and triglyceride were significantly 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 adipocytes. 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.