My current problem is that my cells die when I grow them. It is an MC38 cell. The medium I use is RPMI modified, without calcium nitrate, with 2.05 mm l-glutamine. I add 10% FBS and 1% penecilin/streptomicin. I've tried to revise my technique many times, but, I always get the same result. I would like to have your help or advice in this case to be able to continue my work.
Hello Kodzo Prosper Adzavon,
I feel you should supplement your culture media with 50µM beta-mercaptoethanol when you add the media to the cells. Beta-mercaptoethanol helps to stabilize certain components in the media such as L-glutamine as well as reduce oxidative stress on the cells.
Due to the presence of anaerobes in the colon there is decrease environmental oxygen, and therefore the colon becomes highly hypoxic compared with most of the other tissues. Hypoxia causes decrease in ATP synthesis and increase in ROS levels at the same time as a result of decrease in the activity of the cellular antioxidant system, which may lead to oxidative stress.
Beta-mercaptoethanol is a potent reducing agent used in cell culture medium to prevent toxic levels of oxygen radicals.
Maybe the presence of free radicals in culture could be causing cell death. While some cell lines are able to combat the oxidative stress on their own, in other cases like yours, the cells may not. Give it a try.
Best.
do the cell line really need lack of calcium for growth?
https://www.kerafast.com/productgroup/665/mc-38-cell-line
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