I am using HEK 293T cells, just want to know if we can neutralization the HEK cell line with DMEM growth medium and put in cell culture flask or centrifuge before putting in cell culture flask.
As long as you are diluting the trypsin-EDTA adequately with complete cell culture medium (at least 1:3 or so), you don't need to centrifuge the cells.
Hello Akbar Abdul Rasool Kanji
As soon as the HEK 293T cells have detached, add some complete DMEM growth media containing FBS to the flask (the FBS present in DMEM will inactivate trypsin).
Using this cell suspension, pipette required volume of cells into a new flask at required split ratio. Leave cells overnight to recover and settle. Then you may change the growth media to get rid of any residual trypsin.
Best.
It is not clear what you mean by "neutralization" of HEK 293T cells in this context. If you are referring to removing trypsin or another cell dissociation reagent used to detach the cells from the culture dish or plate, you can neutralize the trypsin or dissociation reagent by adding an equal volume of complete growth medium, such as DMEM, to the cell suspension.
After neutralizing the trypsin, you can transfer the cell suspension to a centrifuge tube and spin the cells down at a low speed, such as 200 x g for 5 minutes, to pellet the cells. Then you can carefully remove the supernatant and resuspend the cell pellet in fresh growth medium, such as DMEM, before transferring the cells to a new culture flask.
It is important to maintain sterile conditions throughout this process to avoid contamination of the cells. Additionally, be sure to follow standard laboratory safety procedures and protocols for handling cells and cell culture media.
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