Monday, October 9, 2023

The Effects of Cell Culture Conditions on the Proteome and Transcriptome!


 If you've seen me speak anywhere recently, you've probably heard more f-bombs when talking about single cell proteomics than maybe if you heard me last year. Our field is going great, but there are a lot of things on the basic level that complicate even the most basic type of research. 

Here is an example - 

This is a really cool resource where a vendor has compiled 7 cell lines derived from patients with the same disease and these cells were selected because the mutations driving the original cancer are different. 

Super cool, right? Particularly if you are using a TOF for multiplexing and your maximum number of samples you can plex (after carrier and 2 blanks) is 7. 

Imagine that you are sort of dumb and you think that "wow! what a great thing, I can do a proteomics (and/or single cell proteomics) study to investigate what effects the mutational status of each of the cell lines has on the proteome! 

Maybe you'll spend $4k on this kit and go to start culturing these cells to find that EVERY ONE OF THEM REQUIRES A DIFFERENT GROWTH MEDIA????




3 shown as an example. One needs something called "McCoy" + 10% baby cow blood extract and the next one needs RPMI-1640 and the same amount of baby cow blood and the third one needs RPMI + 15% cow serum, but also insulin. 

For someone who basically learned cell culture from YouTube and asking some friends and students really dumb questions, what do you do next? 

More importantly, how do you tell that the changes you're seeing are due to the genetics of your cell lines versus what are just the fact you put insulin in one cell line? 

What things would you worry that would change? 



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