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[A convenient and time-saving method for primary culture of mature white adipocytes from mice]

Objective: To establish a simple, low-cost and time-saving method for primary culture of mature white adipocytes from mice.

Methods: Mature white adipocytes were isolated from the epididymis and perirenal area of mice for primary culture using a modified mature adipocyte culture method or the ceiling culture method. The morphology of the cultured mature adipocytes was observed using Oil Red O staining, and the cell viability was assessed with CCK8 method. The expression of PPARγ protein in the cells was detected with Western blotting, and the mRNA expressions of CD36, FAS, CPT1A and FABP4 were detected using RT-qPCR.

Results: Oil Red O staining showed a good and uniform morphology of the adipocytes in primary culture using the modified culture method, while the cells cultured using the ceiling culture method exhibited obvious morphological changes. CCK8 assay showed no significant difference in cell viability between freshly isolated mature white adipocytes and the cells obtained with the modified culture method. Western blotting showed that the freshly isolated adipocytes and the cells cultured for 72 h did not differ significantly in the expression levels of PPARγ protein (P=0.759), which was significantly lowered in response to treatment with GW9662 (P < 0.001). GW9662 treatment of the cells upregulated mRNA expressions of CD36 (P < 0.001) and CPT1A (P=0.003) and down-regulated those of FAS (P=0.001) and FABP4 (P < 0.001).

Conclusion: We established a convenient and time-saving method for primary culture mature white adipocytes from mice to facilitate further functional studies of mature adipocytes.

Comments：

The study has successfully established a simple and low-cost method for primary culture of mature white adipocytes from mice. The modified mature adipocyte culture method proved to be effective in maintaining the morphology of the cultured adipocytes and exhibited no significant difference in cell viability compared to freshly isolated adipocytes.

Furthermore, the study successfully detected PPARγ protein expression and mRNA expression levels of CD36, FAS, CPT1A, and FABP4 in the cultured adipocytes, indicating the potential for functional studies.

The study has provided a valuable contribution to the field of adipocyte research, and the established method could be applied to further understand the metabolic functions of mature adipocytes in mice.

Related Products

Cat.No. Product Name Information
S2915 GW9662 GW9662 is a selective PPAR antagonist for PPARγ with IC50 of 3.3 nM in a cell-free assay, with at least 10- to 600-fold functional selectivity in cells with PPARγ versus PPARα and PPARδ.

Related Targets

PPAR