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Our lab tried your protocol 3 times. Each time, 90% of the cells...

Barbara Summers Nov 21, 2014

Protocol: Isolation and Culture of Bone Marrow-derived Mast Cells

Our lab tried your protocol 3 times. Each time, 90% of the cells adhered to the flask. We had to hit the flask very hard to get cells in suspension. When replated, the cells stuck again. Any idea why we saw mostly adherent cells? After the first 5 days, will most of the cells be adherent?

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Sandra M. O’Neill Author Answered Nov 25, 2014

Biotechnology Department, Dublin City University, Ireland

When feeding the cells it is important to take only the non-adherent cells. The cells adhered to the flask are not mast cells, so try not to get them in suspension. The mast cells yield in the begining is very low, but it will be 30-40 million by passage 9, and there will be less and less adherent cells. We also use flasks for non-adherent cells (cat.no. in the protocol).

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