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An In Vitro Adipocyte Model For Evaluating The Effects Of Environmental Obesogens
Citation:
Watkins, A., C. Wood, AND B. Abbott. An In Vitro Adipocyte Model For Evaluating The Effects Of Environmental Obesogens. Presented at 52nd Annual Teratology Society Annual Meeting, June 23 - 27, 2012.
Impact/Purpose:
to be presented at the Teratology Society Meeting, Baltimore, MD, Jun 23-27-2012
Description:
Obesity is a growing concern in the US population. Current interest is high in the role played by environmental factors called obesogens that may contribute to obesity through developmental exposure. Given the importance of understanding the role that these factors play in lipid homeostasis, we are developing an in vitro model to study the effects of various potential obesogens on adipogenesis and lipid accumulation. Pre-adipocyte 3T3-Ll cells (Zen-Bio, RTP, NC) are mouse fibroblasts that differentiate into adipocyte-like cells and accumulate large lipid droplets in response to medium with high serum content. We have used these cells to evaluate the effects of potential obesogens. Progress to date includes taking the cell line through the differentiation process and staining the intracellular droplets with Oil Red 0 to confirm lipid accumulation. A spectrophotometric method based on Oil Red 0 content is under evaluation as a quantitative measurement of lipid accumulation in the cells. Additionally, work is underway to develop morphometric parameters for measuring changes in lipid droplet characteristics and abundance. Nuclei have been stained with Mayer's hematoxylin in order to quantify the number of cells per unit area. Pilot studies suggest that administration of a potential obesogen, perfluorooctanoic acid (PFOA), to 3T3-Ll cells results in morphological changes and increased lipid accumulation in the cells. RNA has been extracted from treated, differentiated cells for analysis of gene expression. Preliminary experiments using quantitative PCR suggest that genes associated with lipid metabolism and storage are altered in response to PFOA exposure (1-200IlM). Additional candidate genes to confirm these preliminary findings are included in our studies. Our early experiments demonstrate that the 3T3-Ll system is a viable model for studying induction of lipid accumulation in cells and may also be useful for evaluating effects on adipocyte differentiation. Thus far o