Vanderbilt Summer Science Academy

Cell Line Proliferation Patterns of Glioblastoma p53 Controls—Human Non-Small Lung Carcinoma Cell Line (H1299) and Human Osteosarcoma Cell Line (U2OS)—Determined using Doubling Time

This study focuses on H1299 and U2OS proliferation using doubling time—the time (days) required for a cell population to double. Doubling time is appropriate because cell proliferation with optimal conditions of media and temperature follows exponential growth. H1299 and U2OS are controls for p53 binding: H1299 lacks p53 and therefore is a negative control and U2OS overexpresses p53 and is a positive control. These cell lines will be used as controls to test binding specificity of antibodies in glioblastoma. Subpopulations of glioblastoma cells will be characterized through flow cytometry—the future direction of our experiment. Cells were counted at different times throughout their growth cycle through passaging. This process involves lifting adhesive cells with 0.25% Trypsin, resuspending cells in new media, spinning down the cells into a pellet, and counting the cells using Trypan Blue stain to determine intact—non-colored—cells. Passaging time varied from two to four days after plating. H1299 has a doubling time of 3.47 days and U2OS has a doubling time of 3.20 days (on a 100mm cell culture dish). H1229 has a larger standard deviation (±1.98) than that of the U2OS cell line (±0.57). The results indicate the U2OS cell line grows faster than the H1299 cell line but further investigation is necessary to minimize deviation. This study will execute a single-cell approach, rather than a homogenate analysis, to analyze individual cells in a glioblastoma tumor against the controls. The growth patterns of H1299 and U2OS serve as benchmarks for p53 activity.