I recently did a single cell sort with MCF-7 cells to create my CRISPR cell line. I sorted single cells in a 96-well plate in MEM media with 10% FBS, 10ug/mL insulin, and 1mM sodium pyruvate. It has been a week since the sort and I cannot see any cells yet. I understand that single cells would take sometime to grow into colonies, but I was also wondering if anyone has any helpful suggestions on improving cell recovery? I have also read about conditioned media. At what point should I think about changing the media?
I appreciate any and all tips and suggestions!
Thank you.
I would not plan to switch out the media as removal of it can take your cells with it if you are not extremely cautious.
I have had luck in difficult to grow cell lines by increasing the FBS to 40% final and just replacing with 100 ul of media every few days as it evaporates off.
At one week i would think you should find at least some wells with a cell or more in them if the cells survived the sorting fine.
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