Effects Of Creatine Supplementation On The Viability Of Walker 256 Mammary Carcinomas

Doxorubicin (DOX) is a potent chemotherapeutic agent with significant cytotoxic effects. Recent evidence has shown that creatine (Cr) can significantly reduce the degree of oxidative damage caused by DOX and promote the survival of non-cancerous tissues. However, it is unknown if Cr treatment would minimize the cytotoxic effects of DOX. PURPOSE: To determine if supplementation with Cr alters cell viability in a tumor cell line when combined with DOX over a 12 hr time period. METHODS: Walker 256 mammary carcinoma cells were cultured in growth medium (90% DMEM 10% FBS) until they reached 90-95% confluency. Cells were seeded onto a 96-well plate at a density of 10,000 cells/ml with fresh growth media and incubated for 24 hr prior to experimentation. Cells were then exposed to growth media containing either 10 μM of DOX, 10 mM of Cr, 10 μM DOX + 10 mM Cr, or regular growth media as a control for an additional 12 hr. Cell viability was assessed at 0 and 12 hr using an EarlyTox™ Cell Integrity Kit and analyzed via a Nikon live cell confocal imaging system. RESULTS: At 12 hr post-treatment, DOX had significantly lower cellular viability (18.25 ± 3.4% non-viable, P < 0.05) compared to control (0.46 ± 0.70% non-viable). No significant changes in viability were found between the Cr (0.75 ± 0.25% non-viable) and control group. Cr + DOX had significantly improved cell viability (10.21 ± 2.31% non-viable) compared to DOX. CONCLUSION: Cr appears to provide a protective effect against the cytotoxic effects of DOX.