Hello,
I am conducting MTT assays on various cancer cell lines. While I have no problems with any of the other lines, I have seen that a good number of the SH-SY5Y cells I plate for the assay (20000 cells per well in a 96-well plate) become detached when I change the medium to initiate treatment (at 24 h post-plating). I plate the cells in medium containing 10% FBS and treat them 24 h later in medium that does not contain any serum. Could anybody help me with this problem? Thank you very much!
I have experienced similar case when I cultured SiMa cell line.
So, I wrote comment expecting my expirience to be your help.
Neuroblastoma cell line have weak attachment.
Therefore this cell line is easier to detach by shear stress of fluids.
In my case, delicate pipetting/aspriration help the more cell ramain to attach.
And using a coating material such as poly-D-Lysine may help your cell to be adherent.
Thank you very much, Beomju. Unfortunately, I tried to be extremely careful with my pipetting and I also tried PLL-coating, but this did not improve my results. Thank you.
Dear Beatriz Duran ,
Did you try Matrigel (1:50 ratio) to attachment of SH-SY5Y cells?
Dear Simge,
Thank you very much for your recommendation. I did not try, as it would not be worth conducting these experiments in this manner. Thank you!
I successfully grow these cells on 96-well plates with a glass coverslip bottom coated with 50-100 ug/ml PDL. To minimize cell detachment I change only 50% of the media during media change or drug treatment. They do tend to clump and pull up after 3 days when not differentiated. Best of luck to you.
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