hi all,


i am new to cell culture experiments and i have standarised the MTT assay for the HepG2 cell line.


now i want to standrize the same protocol for 2 cancerous cell line
--------- one is RAJI cells
---------and other is K562 cells



now i am litttle bit confused about it that is it possible to use the MTT for the suspension culture,

does any body have idea for the suspension culture regarding this.



please do the needfull



thanx in advance..

It is easily possible to do the assay with suspension cells. One of the most widely used screening methods for HIV-1 inhibitors is based on the fact that HIV-infected cells have cytopathic effects whereas non-infected cells survive. The assay is performed on MT-4 cells (a T-cell line, suspension).

I used to do it in 96 well plates, so I can centrifuge the cells.
and the procedure was the same than for adherent cells in 96 well plates.

You can also use the soluble MTS reagent, in which case no centrifugation is necessary.

I've been involved in culturing primary lymphocytes and using MTT assay to evaluate drug sensitivity towards the cells. The standard MTT protocol is the same for both adherent & suspension.

I used v-bottom 96-well microtiter plates (Greiner) as to pellet the suspension cells after MTT treatment. The results are consistent compared to the flat-bottom ones..............& dissolved the cells in DMSO before analyzing it.

Good luck !.................