We are trying to passage MCF10A cells but we encounter some difficulties. We are using 0.25% tripsin and we are cultivating them with MEBM supplemented accordingly to manufacturers (LONZA). Any suggestions? What do you think we are doing wrong? Thanks a lot.
Zhilin Yang
0.25% is too high. use either 0.05% (ATCC recommended) or 0.025% (lonza recommended).
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Linglan Fang
Zhilin Yang How long does it usually take for MCF10A to detach from the flask at 37 ˚C with 0.05% Trypsin? Thanks!
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Zhilin Yang
Linglan Fang If you pre-wash with either PBS or a small amount of Trypsin , it takes ~10-20 min to detach the cells
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