This question is part of the following Ask The Expert session:
I guess the first question is…were the cells growing in the lab of the collaborator and did you switch the growth method. Use a shaker flask without baffles. Also the best thing to do in the future is to split the cells into your media instead of pelleting them. I try not to pellet cells if possible. I would not try and continue growing those cells that you have. Get more cells from the collaborator. Let say you get 50 mls of culture. I would let the 50 mls in a sterile falcon tube or equivalent sit for a while to allow the cells to settle. Then remove the medial above the cells but leave some for safety and conditioning. Then re-suspend those cells and the little bit of media they are in in some volume of fresh media. Do not let your cells get below 500,000/ml for the first several passages. I hope this helps. I suspect the centrifugation was the problem?