Identification of suitable methods to determine cell viability

Camilla Habo, Stine Hesselholt Sloth, Ida Johanne Kyhnauv & Grace Gloria Namatovu

Studenteropgave: Bachelorprojekt

Abstrakt

To measure cell viability there are a broad range of cell based assays to use. The in vitro methods differ in relation to how they respond to the cells, and by what mechanism they measure them. In this study we will examine how these differences are reflected in the colon cancer cell line DLD-1 measured with different in vitro assays: MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazoliumbromide), Alamar Blue, and cell counting, respectively. The DLD-1 cells were treated with the polyphenols Resveratrol and Pterostilbene, that known for their decreasing effect on cell proliferation are examined in relation to cancer treatment. We treated the cells with 3.75, 7.5, 15, 30 or 60 uM for 48 hours. The cell viability was examined by the methods described above. The results show that cells treated with resveratrol and pterostilbene when measured with Coulter counter had an IC50 value of 28,6 µM and 20,4 µM, respectively. When measured with MTT the IC50 value was much higher at above >60 µM for both polyphenols. As for Alamar Blue the IC50 values was also higher than when measured with Coulter Counter, at over 60 µM for resveratrol treated cells and 55µM for pterostilbene. The higher IC50 values are in this report presumed to be due to the staining methods of both Alamar blue and MTT. For the DLD-1 cells treated with resveratrol and pterostilbene the cell diameter increases from around 15um to 25um and 20um respectively. For both staining methods the color compound are reduced in the mitochondria, with a larger cell diameter, we propose that this gives an overestimate of the compounds effect on decreasing the cell proliferation.

UddannelserMedicinalbiologi, (Bachelor/kandidatuddannelse) BachelorMolekylærbiologi, (Bachelor/kandidatuddannelse) BachelorKemi, (Bachelor/kandidatuddannelse) Bachelor
SprogEngelsk
Udgivelsesdato29 maj 2017
Antal sider38
VejledereOle Vang

Emneord

  • MTT, Alamar Blue, Coulter Counter, DLD-1