(188al) Differentiated Transcriptome Profile of 3T3-L1 Adipocytes in 3-D in Vitro Culture | AIChE

(188al) Differentiated Transcriptome Profile of 3T3-L1 Adipocytes in 3-D in Vitro Culture


Janorkar, A. - Presenter, University of Mississippi Medical Center
Turner, P., Univesrity of Mississippi Medical Center
Garrett, M., University of Mississippi Medical Center
Didion, S., University of Mississippi Medical Center
In vitro models that capture in vivo cellular morphology and functionality offer a relatively straightforward way to understand key effects of dietary fatty acid types and dosages as well as cytokines on cellular metabolic outcomes. Previously we have demonstrated: (1) enhanced adipogenic differentiation in 3-D spheroids compared to the traditional two-dimensional (2-D) monolayer cultures based on triglyceride accumulation, CD-36 and CD-40 protein expression, and peroxisome proliferator-activated receptor-gamma (PPAR-gamma) and adiponectin mRNA expression; and (2) LA-fed cultures have reduced metabolic function and enhanced lipolysis in response to TNF-alpha. In the current study, we present an in-depth genomic analysis (whole transcriptome analysis) of the 3-D spheroid culture compared to the 2-D monolayer culture of 3T3-L1 preadipocytes as they are differentiated using adipogenic cocktail, matured in presence of LA, and subsequently exposed to TNF-alpha. Additionally, expression of key adipogenic genes in 2-D monolayer and 3-D spheroid cultures were compared to the adipose tissue isolated from mouse perirenal fat deposit to identify which culture method more closely resembled the isolated adipose. We also present transcriptome level evidence that the 3-D spheroids, with their reduced cell-substrate interactions, suppress Rho-ROCK signaling pathway to enhance PPAR-gamma expression and achieve enhanced adipogenesis. Overall, we have demonstrated the feasibility of genome-wide analysis of our in vitro 3-D spheroid model and its superiority over the conventional 2-D monolayer model.