Differentiation of 3T3‐L1 preadipocytes is inhibited under a modified ceiling culture

Abstract

Ceiling culture is an inverted and closed cell culture system which represents a novel method for exploring adipocyte characteristics and function. Although the role of ceiling culture in mature adipocyte dedifferentiation has been extensively studied, its potential effects on preadipocyte differentiation remain unclear. In this study, we established a simplified dish ceiling culture method for 3T3-L1 preadipocytes and showed that our novel ceiling culture method could reproduce the function of the traditional flask ceiling culture. Then, we investigated the effects of ceiling culture on 3T3-L1 preadipocyte differentiation by Oil red O staining and RT-qPCR. The results showed that ceiling culture significantly impaired triglyceride accumulation and adipogenic marker genes expression in 3T3-L1 preadipocytes. These findings suggest that ceiling culture inhibited 3T3-L1 preadipocyte differentiation while inducing mature adipocytes dedifferentiation. Taken together, our data facilitate the understanding of the property of ceiling culture and promote the study of revealing the underlying mechanisms of mature adipocytes dedifferenatiation.

An optimized method for Oil Red O staining with the salicylic acid ethanol solution

“…After cell adhesion, the medium was changed into DMEM/F12 complete medium containing 100 μmol/L sodium oleate for 48 hours. Primary adipocytes were extracted from fat tissue of 5-week old rat and cultured for three days by using a modified ceiling culture method [ 34 ], the slide with adherent adipocytes was put in 12-well plated and fixed for staining. The above cells will be used randomly for subsequent staining.…”