Dear all,
I am doing some CRISPR experiments in neonatal fibroblasts.
After a lot of optimization, the system works really well in these cells. As you will know, fibroblasts don´t really like to grow alone :) So, my single cell clones die within a few days. I already tried to use conditioned medium, but with no success. I guess these cells need to have a cell-cell contact to grow. For this purpose, I am going to use feeder cells on which I would like to cultivate my target cells. I would like to use the same cell-line as my feeder cells. MMC-treatment of these cells works really well. My question is: How should I separate my feeder cells from my target-cells? The only idea I have is to do it by serial dilutions. But this would result in very high passage numbers. Do you may have other suggestions?
Best regards
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