Effects of phosvitin on intestinal epithelial cell viability analyzed by flow cytometry and Alamar blue cell viability assay

Phosvitin is a phosphoprotein which makes up 7% of the proteins of chicken egg‐yolk, has been shown to have a wide variety of properties including cytotoxic and antigenotoxic effects on selective cancer cell lines. The effects of phosvitin on normal cell line is not yet known. Therefore, diploid, normal rat intestinal epithelial cells (IEC‐6) were used to investigate the effects of phosvitin on cellular health and viability. The focus of this research project is to analyze cytotoxic and antigenotoxic effects of phosvitin at varying concentrations in cell culture media. IEC‐6 cells were grown on plastic plates using DMEM media and 10 % CO 2 in a humidified incubator following standard protocol. Alamar blue cell viability assay and flow cytometry were used to determine the effects of phosvitin on internal complexity of the cells. Data gathered from the flow cytometer involved the use of propidium iodide which allows for cells in suspension to be quantified based on forward scatter. Data is plotted based on propidium iodide versus forward scatter to determine cell count and health. Preliminary flow cytometry data were employed to plot forward scatter versus side scatter using propidium iodide as an indicator for cells treated with serial dilutions of phosvitin. These data suggested that phosvitin has cytotoxic effects on IEC‐6 cells, decreased number of healthy cells in cell suspension in a dose dependent manner. These findings were confirmed using Alamar blue cell viability assay to determine the viability and apoptosis of IEC‐6 cells. We conclude that phosvitin can be used as a cytotoxic agent in different disease conditions including to control cancerous growth.