High throughput screening of small molecules for enhancing adipogenesis in vitro

Abstract

Adipose tissues have been known to be the major site for energy storage and metabolic maintenance. Despite being researched for many decades, expanding and culturing primary adipocytes has remained difficult. To circumvent this problem, many labs in the field are using adipocytes differentiated from induced pluripotent stem cell-derived mesenchymal stem cells (iPS-MSCs), which can expand much more easily. However, these iPS-MSC-adipocytes are typically very immature, with only low levels of lipogenesis. In this study, we utilised small molecule high throughput screening (HTS) methods to promote the adipogenic differentiation and maturation of iPS-MSCs. We found that a small molecule combination 2+DO was able to significantly promote and enhance adipogenesis. We also optimized the iPS-MSC seeding density and glucose concentration to optimize subsequent adipogenesis. Finally, we tested our small molecule combination 2+DO on primary pre-adipocytes as well. RNA profiling by qRT- PCR showed that many adipogenic biomarkers such as LPL, SREBF1, FGF2, IRS2 and PPARG were significantly upregulated by 2+DO, as compared to normal adipogenic media. Taken together, we have demonstrated that HTS is a viable method for screening novel small molecules that enhance adipogenesis. This platform will provide a strong foundation to model fat-related metabolic diseases in vitro in future.