To prevent loss of myoblastic cells, cultures should be subcultured before they become confluent. During subculturing, to detach the cell from the flask/dish and to avoid cell to cell adhesion usually trypsin and EDTA mixture are widely used. And their concentration depends on cell lines to cell lines.
I am going to start working with h9c2 cell lines. According to ATCC (product sheet H9c2(21) (ATCC® CRL1446™ ), I have to use 0.25% (w/v) Trypsin 0.53 mM EDTA solution for detaching the cells during subculturing. So, now I need a protocol to prepare this mixture. Thanks in advance.
Simply order the solution by thermo scientific ;-)
https://www.thermofisher.com/order/catalog/product/25200072
good luck
Mol wt. of EDTA is 372.24 g/mol
First we want to prepare 500 ml (e.g.) of 0.53 mM EDTA solution.
Weight 0.099 g of EDTA and prepare 500 ml of solution (0.53 mM)
For 0.25% (w/v) Trypsin of 500 ml, weight 1.25 g trypsin and volume it up to 500 ml with the prepared 0.53 mM EDTA solution.
Dear Mohammad Murshedul Alam, Could you tell me which solvent should I use in in this case?
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