Meeting Abstract
P2.99 Sunday, Jan. 5 15:30 Using Molecular Markers to Rapidly Characterize Adipogenic Stem Cell Differentiation PATTON, M.P*; TEMKIN, A.; ELLISOR, D.; KOHNO, S.; GUILLETTE, L.J., ; SPYROPOULOS, D.D.; College of Saint Benedict & Saint John’s University, St. Joseph, MN and Medical University of South Carolina (MUSC), Charleston ; MUSC; MUSC; MUSC; MUSC; MUSC michaelapatton6491@gmail.com
Research demonstrates that embryonic exposure to contaminants considered obesogens may cause a predispostion to obesity. The obesogenic potential of crude oil and dispersant from the 2010 Gulf of Mexico Deepwater Horizon incident is of particular interest due to large numbers of exposed marine organisms and humans. We are developing a novel method of investigating adipogenic induction in stem cells exposed to oil/dispersant mixtures. Cell type-specific molecular markers were selected to characterize the adipogenic differentiation pathway. Human mesenchymal stem cells (hMSCs) were cultured under adipogenic conditions. RNA was isolated from untreated hMSCs, preadipocytes and cells at day 15 of adipogenic differentiation. Quantitative real time PCR showed hMSCs highly express specific markers CD166, CD105, CD90 and CD44, with CD105 being the most stringent MSC-specific marker. Differentiating stem cells showed increasing expression of adipocyte markers FABP4, LEP and ADIPOQ and decreasing expression of MSC markers and the preadipocyte marker FSTL1. To address the potential obesogenic effects of the oil/dispersant mix in an ecologically-relevant model organism, American alligator stem cells were generated from stage 19 embryos. Adipogenic induction in these cells was demonstrated via Oil Red O staining, and adipocyte-specific genetic markers are currently being developed for alligator cells. Future studies will utilize molecular markers to determine the obesogenic potential of the oil/dispersant mix on human, alligator and pygmy sperm whale stem cells in more rapid, quantitative manner than current techniques, such as triglyceride quantitation.
