I am looking to stockpile some conditioned medium prior to starting a clonal isolation of HepG2 cells. It has been my previous experience that most HepG2 cells sorted as individual cells into wells of a 96-well plate did not survive. Though this could have been due to other circumstances created by my methods, those cells that did survive grew soooo slowly. I am looking to improve upon either or both survival and initial growth rate. One of the ways I am looking to do this is sorting directly into conditioned medium and maintaining the cells in conditioned medium during the early clonal expansion.
So now that you have an idea what I want this conditioned medium for, my question is: How long can I store conditioned medium and still expect it to serve my purpose? Should I store frozen at -20 C, -80 C, or would 4 C be sufficient?
I would store it at -80 degree C. When ready to use thaw it quickly by incubating at 37 degree C and shaking the plate in a shaker.
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