I am currently culturing some HEK cells with absolutely no issue, they are being cultured with the standard DMEM + 10% FBS media.
For a particular experiment, I need to introduce exogenous insulin for a variety of different time points and I am noticing that my cells are dying within 5-10 min of insulin addition.
I am adding insulin in at a concentration of 10ug/ml. The cells are grown in a 6 well and all those that dont have insulin added to them are surviving but the addition of insulin is resulting in all the cells lifting within 5 minutes. If someone could provide some guidance.
Smarth Narula Perhaps you can try using serum free DMEM media while you are treating and you can also try using less concentration of insulin (1-5 ug/ml).
Dear Smarth!
Plese You look a t the article
https://academic.oup.com/endo/article/154/1/375/2423596
Perhaps insulin through its receptors on HEK293 cells enhances the accumulation of APP, which is toxic to these cells
I think you should do a range of insulin dilution and use the limit dilution for your experiment
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