I'm doing primary cell culture and I want accurate and more sensitive assay to deteremine the cell proliferation in real time. Since I'm using 3D cultute systems, I want an assay which should not get affected with the use of different culturing scaffolds.
-- Thanks in advance.
Hi, I would suggest CellTrace ro simlar (CFSE; PKH26)dye, you can combine different colors (gree, red violet) so you can have one population marked with one color and follow them (some are suitable for microscopy too, not only flow). It should allow for minimal handling of cells (check different time points and cells in single culture too) and you can combine surface marker at FACS too.
good luck !
We're using CellTrackers (https://www.thermofisher.com/il/en/home/life-science/cell-analysis/cell-tracing-tracking-and-morphology/cell-tracking.html).
The advantages of this method are: low to no cytotoxicity (depends on the cells you're working with), wide range of visualization options (infra-red, fluorescence, confocal, in vivo imaging etc.), high stability of the signal and relatively long time of action (we've succeeded to monitor the cells up to 24-30 days (again, depends on the type and the size of your cells). The signal can be quantified by the intensity and the number of foci.
If not, you could simply sample equal parts of the 3D culture, resuspend the cells in 2D culture and run an old good XTT/MTT assay.
perhaps could you try an indirect method by measuring at various intervalls either glucose consumption and/or lactate release in an aliquot of the culture medium. We have done that in hybridoma cultures and we could follow quite accurately cell proliferation, at least during the exponential growth phase.
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