Problem with HeLa not attaching to six well plates - (Aug/07/2007 )

my box of 6-well plates says costar 3506 not 3516 didn't look up the difference but on the outside my box says tissue culture treated...

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never mind they are the same just individually vs bulk wrapped.

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Thanks becaf22, I ended up throwing away the culture and started a new one (following fred33's suggestion, THANKS fred33!) now the cells are growing nicely again in the plates. I tested for mycoplasma but could not detect it. Could it have been that the cells were too old? I mean, I have passaged them many many times (probably some 80 passages). Thanks to all you guys!

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may be because of the density of cells seeded per well.

try to increase the density of the cells.


May I ask ... did you count the cells in trypan blue before seeding the well?
... how many cells did you seed per well ??
... how long did you trpsinize the cells?

hope this may help.

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Thanks Minnie, yes I count routinely the cells before seeding the well, I typically seed at 100 000 cells/mL (3mL). This had worked all the time before these "obscure" two weeks in which my cells simply decided not to attach. For trypsinization I usually give two to three minutes.

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