The Effects of Oxidative Stress on Inducing Senescence in Human Fibroblasts

摘要

Oxidative stress, specifically from hydrogen peroxide exposure, was performed to determine if it induced senescence in cell lines such as Hela cells and primary human fibroblasts. The purpose of this experiment was to find the optimal stage, concentration, and time of exposure to induce the greatest number of senescent cells. After dividing cultures of both the Hela and human fibroblasts cells in order to reduce confluency, the cells were placed in a six well plate and exposed to hydrogen peroxide for two hours at various concentrations. The plates were checked at twenty-four-hour intervals, and then fixed with senescence associated beta-galactosidase as a biomarker to observe the senescent cells in culture. It was hypothesized that hydrogen peroxide exposure would increase the number of senescent cells due to the accumulation of reactive oxygen species. The data indicated that, there was an increase in the number of senescent cells following 48 hours of treatment. The number of senescent cells peaked following 72 hours of treatment and did not change significantly as result of 96 hours of treatment supporting our hypothesis.