Purpose: In this study, we determined that the troglitazones could induce uniform adipogenesis of human mesenchymal stem cells (MSCs) within a short time in a dose- and a time-dependent manners.
Materials and Methods: Human MSCs were isolated from bone marrow and cultured in basal or adipogenic medium in the presence of 0 50 μM troglitazone for 5 days. Then we performed flow cytometry, RT-PCR and western blot analysis.
Results: In FACS assay, troglitazone induced adipocyte differentiation in a dose-dependent manner. At concentration of 25 μM troglitazone in adipogenic medium, over 50% of the cells differentiated into adipocytes at day 5. This was accompanied by increased mRNA levels for the adipocyte gene markers (LPL, aP2 and PPARγ) in RT-PCR. In western blot analysis, we found that ERK phosphorylation was inhibited in the early stage of adipogenesis.
Conclusion: Through the addition of troglitazone as a PPAR γ agonist, we could get the uniform adipogenic differentiation within a short time. Thus, troglitazone directly regulates differentiation of human MSCs into adipocytes; induced PPAR γ expression may play a key regulatory role in this process. And we suggest a role for ERK as a regulatory switch for these differentiation pathways