A CYTOMORPHOLOGIC CHARACTERIZATION OF PANC-1 CELLS EXPOSED TO INCREASING GLUCOSE CONCENTRATIONS USING REPEATED MEASURES ANALYSIS TECHNIQUES

Abstract

Previous studies in our laboratory provided evidence that PANC-1 cells can be manipulated into insulin-producing cells by altering the culture medium with increasing amounts of glucose. However, there is a paucity in the literature regarding cytomorphologic characteristics of PANC-1 cells under normal culture conditions and when challenged with increased amounts of glucose. This experiment evaluated the cellular, nuclear and cytoplasmic characteristics within groups of PANC-1 cells at 24-, 48-, and 72-hours exposed to varying percentages of glucose in media. Initially, cells were grown in flasks with a control medium and then split into four separate groups containing control media or media containing an extra 1%, 2.5%, or 5% extra glucose. Cells from the three cultures were plated (1 x 105 cells/well) and treated with control, 1%, 2.5%, or 5% glucose for 24-, 48-, and 72-hours. Cells and supernatants were harvested, and the cytomorphologic characteristics were compared across all three-time phases. Nominal data were analyzed using non-parametric statistics calculating mean ranks to compare changes within all four groups using the Kruskal-Wallace H statistic. There were statistically significant differences in morphological characteristics within glucose concentration groups at 24-, 48-, and 72-hours (p<0.05). The most telling was the stability in ranks observed in the nuclear characteristics at the 2.5% extra glucose concentration. At the 72-hour phase, ranks for the nuclear and other characteristics became more variable, indicating stress and degeneration of the PANC-1 cell populations. This study contributes valuable semi-quantitative data regarding the viability and function of PANC-1 cells over 72-hours with increasing glucose challenge and demonstrates proof of concept that PANC-1 or other insulin-producing cell-lines could be further engineered into useful components in drug delivery and organ-function replacement applications.