Introduction Rodent insulinoma cell lines, such as βTC3 cell, are effective tools to understand β cell function and acquire clues to obtain a continuous.

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Introduction Rodent insulinoma cell lines, such as βTC3 cell, are effective tools to understand β cell function and acquire clues to obtain a continuous supply of insulin producing β- cells (Cozar-Castellano, 2008). Due to the high demand and low supply of human islets of Langerhans for transplants, these cells provide a trail run of potential proliferation techniques of islets before actually testing the on human islets. Hypoxia is defined as an inadequate supply of oxygen reaching tissues, organs or cells. Under normoxic conditions hypoxia proteins are not active but under hypoxic conditions, hypoxia inducible factors (HIF) activate, mainly HIF-1α and HIF-2α, regulating homeostasis in the human body. HIF-2, one of the members of the HIF family related to hypoxia, but only present in certain endothelial and fibroblast tissues (Loboda, 2010). We hypothesize that βTC3 cells placed under hypoxic conditions will present a greater expression of HIF 2 α the longer the hypoxic time exposure. Methods Results Figure 1 A. Hypoxic group of BTC3 cells at t=0 hrs 10x magnification, B. Normoxic group of BTC3 cells at t=0 hrs 10x magnification, C. Hypoxic group of BTC3 cells at t=96 hrs 20x magnification, Normoxic group of BTC3 cells at t=96 hrs 20x magnification Figure 3 Relative values of average protein quantification obtained from western blot Image J and Excell analysis from normoxic and hypoxic groups. Conclusions T-test results implies no statistical difference among the normoxic and hypoxic groups but a clear distinction is viewed along the pictures (Figure 1). To further identify whether HIF 2α was expressed and a statistical difference was obtained, βTC3 cells were exposed to 96 hours of hypoxic conditions based on a publication stating the greater the time of hypoxic exposure the greater the expression of HIF 2α (Holmquist-Mengelbier, 2006).The presence of HIF 2α is more vividly seen the more time βTC3 cells are exposed to hypoxic conditions. Future experiments need to be conducted regarding more sensitive cells or cell lines such as MIN6 cell lines and insulin producing islets of Langerhans. MIN6 cell lines exhibit similar characteristics to islets and are also more sensitive and more difficult to cultivate providing much better comparison to real islet structure and physiology (Ishihara H, 1993). Acknowledgments Special thanks to NIGMS/HHMI funded Bridge to Baccalaureate Program for providing me the opportunity and resources to complete this project. Thanks to workers and volunteers of the Diabetes Research Institute Miller School of Medicine, in particular Julio Aleman, Marta Garcia and Dr. Nancy Vargas for providing me the assistance and materials for this project. Thanks to Silvia- Cubela Alvarez and Dr. Juan Dominuguez-Bendala for accepting me into the Diabetes Research Institute and providing me the assistance, and guidance to completing this project. Cells Cultured Hypoxia Normoxia Protein Extraction HIF -2 α Western Blot Activation of Hypoxia Inducible Proteins in βTC3 Cells Abelardo D. Montalvo, Julio Aleman, Marta Garcia, Nancy Vargas, Silvia Alvarez-Cubela, Dr. Juan Dominguez-Bendala Miami Dade College, University of Miami