I need to harvest the secretome of mesenchymal stem cells. In order to do that, I used serum free medi and 75T culture flask but the concentration was low and not enough to run it on the array chips.
Does anyone have experience?
What do you recommend?
You could concentrate your media do your protein chip. I used centricon device to do the concentration (You could used another provider). I used a small cutoff to keep a maximum of protein.
How long did you cultivate your MSCs in the serum-free media? If you do it too short, they won't have enough time to secrete anything, but if you do it too long, they won't like it either because they need the serum. I would recommend to use serum-containing medium and have a negative control of it on the array. Good luck!
You could try 24H or 36H. No more 72 hours. In fact, you must not wash your cells before deprivation. Just aspirate the proliferation medium and put your collection medium.
Because you concentrate your medium, you could do the collection with large amount of cells, something like 2 T75 or more...
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