I am growing MCF-7 cells and after 4 hours of thawing and putting in a flask, the cells are still floating. I am worried because this is an adherent cell line and should not take this long to attach. Could anyone provide any thoughts on this.
Thanks
Basically, after 4 hours at least some cells should be attached. But just wait a bit longer, typically one over night cycle.
Hi,
Usually after 6-7 hours they should be attached.
Removing any residue of freezing medium from the frozen cells will make them less stressed.
You can do this by thawing the cells in 10ml medium, spin theme down, remove the supernatant, dilute the pellet into fresh medium and then plate the cells
Cheers
Pan.
Hello Vasundhra,
It takes generally overnight for MCF-7 cells to attach properly. I hope stocks are not very old and that might be the main reason that cells are not attaching in expected time. You must be knowing very well about gentle thawing and resuspension. What is generally recommended for MCF-7 is alpha-MEM with US origin FBS. You can add 10ug/ml of insulin for best growth and attachment of these cells. These conditions worked very well for me and a confluent flask was obtained at third day.
Hope it helps, Do let me know if any further assistance required.
Cheers !
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