problem with plating adherent cells in 24-well plates - (Sep/26/2007 )

Hello,

I am currently having a problem with plating my adherent cells in 24-well plates. The cells grow well in flasks, but after I plate them in wells they tend to form clusters and a lot of them die. Can anybody give me advice? Thanks in advance.

dc11

The best way to tell is after you seed the plate, seed the remaining cells to a flask to see if you have the same problem. I guess you somehow over treated cells when you detached cells with trypsin-EDTA.

Hi genehunter-1,

I did seed the remaining cells in a flask and they grew well. I do not know why the cells died when being seeded in plates. The plates work well with another type of cells. Thanks for your reply.

dc11

dc11,

Are the plates you're using Tissue-culture treated, or otherwise treated to make sure the cells can adhere to them?

Hi TheSquire,

Yes, they are. The plates work good with another type of cells I am culturing. Thanks.

dc11


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Do these cells require coating to grow by any chance?

Are the cells clumping in the middle of the well, or all around the well??

Could you have possibly seeded the well with too many cells?? this could explain both the cell death and the clumping......
I have found from working with different cell lines, that the seeding density that works for one cell line does not neccessarily work for another.

Hi Lauralee,

The cells are clumping all around the well. I seeded about 1 X 105 cells each well. Thanks.

dc11

Hi genehunter-1,

The cells do not require special coating to grow. Thanks.

dc11

Maybe gelatin coating can solve your problem? 0.1% RT 1hr, then remove it.