Effect of endogenous methylglyoxal on Chinese hamster ovary cells grown in culture

✍ Scribed by Frank W. R. Chaplen; William E. Fahl; Douglas C. Cameron

• Publisher: Springer
• Year: 1996
• Tongue: English
• Weight: 951 KB
• Volume: 22
• Category: Article
• ISSN: 0920-9069
• DOI: 10.1007/bf00353922

✦ Synopsis

Methylglyoxal is a ketoaldehyde that reacts readily under physiological conditions with biologically relevant ligands, such as amine and sulflaydryl groups. It is produced in mammalian cells primarily as a by-product of glycolysis. The level of glucose, L-glutamine and fetal bovine serum in culture media was found to significantly affect levels of intracellular methylglyoxal in Chinese hamster ovary cells. Medium with 25 mM glucose and 5 mM L-glutamine caused an increase in free methylglyoxal levels of 90 to 100% relative to medium containing 5 mM glucose and 2 mM L-glutamine. Both of these media compositions are representative of those found in commercially available media. Pseudomonasputida glyoxalase I was expressed in Chinese hamster ovary ceils to enhance methylglyoxal detoxification. The Chinese hamster ovary cell clones showed an 80 to 90% decrease in free methylglyoxal levels. The colony-forming ability of these ceils was compared to wild-type Chinese hamster ovary ceils under conditions found to cause elevated methylglyoxal levels. The wild-type cells showed a 10% decrease in colony-forming ability relative to the clones. This decrease was found to be statistically significant (P > 0.99) by analysis of variance. The variation in colony-forming ability amongst the clones was statistically insignificant. More importantly, the clones showed increased colony-forming ability relative to the wild-type cells unde r conditions of higher methylglyoxal production with fair to good statistical significance (P > 0.75 to P > 0.95). This result is the first quantifiable evidence that endogenously produced methylglyoxal can negatively affect cell function under conditions found in animal cell culture.