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Lab_Report_#7 - consequences of chronically high levels of...

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Metabolic Effects of Elevated Glucose On Human Retinal Pigment Epithelia Cells Jaime Figlan Hatham Shawqi Department of Physiology At Michigan State University Tuesday, March 10 th , 2008
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Abstract The purpose of this experiment was to examine the effects of high levels of extracellular glucose on the amount of lactate within hRPE cells. It was hypothesized that high levels of glucose would increase the amount of lactate within the cell. Since high levels of lactate are considered to represent tissue hypoxia and acidosis, a direct relationship between glucose levels and lactate levels would support the idea that elevated levels of glucose are harmful to cell tissues. The 22mM glucose hRPE cell line contained --------------------------
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Introduction Diabetes is a serious immunological disease in which normal control of circulating glucose levels is lost. There are many negative consequences of failure to successfully regulate glucose levels. In this experiment, we will examine the
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Unformatted text preview: consequences of chronically high levels of glucose concentration on human retinal pigment epithelial cells by comparing hRPE cells with glucose concentrations of 5.5mM and 22mM glucose. The hypothesis is that elevating levels of extracellular glucose from normal will increase the production of lactic acid. An accumulation of lactate within the cells is considered to represent tissue hypoxia, and therefore we will use the cellular production of lactate as the dependent variable. Therefore, we predict that the 22mM glucose cell culture will have a greater amount of lactate within the cell. Methods and Materials The 5.5mM and 22mM hRPE cells were prepared following the instructions in the Lab Manual (Hootman). The spectrophotometer was then used to determine the amount of lactate in the samples, as well as the amount of protein in the cell, using the assays described in the Lab Manual (Hootman)....
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