Dear All,
i was studing the role of one cytokine in promoting cell proliferation, performing FACS analisis i got this results;
Average of cells in phase G1/G0 30%, S 17%, G2/M 21% in untreated cells
and
G1/G0 27%, S 15%, G2/M 24% in treated cells.
Comparing both groups, only data from G2/M in treated cells are statistical significant .
I was wondering if we can conclude that this cytokine promotes cell proliferation since only the G2/M phase ( but not S ) is statistical significant in both groups.
Thank you in advance.
Giovanni
Hi Giovanni, according to your data, there seems no big difference between the treatment group and control group. To verify these data, you can follow several ways:
1. You probably need to do cytokinetic analysis of cell cycle distribution: you can synchronize your cells at G0/G1 phase by starvation, for example, to starve you cells in medium w/o serum for 24 h; And then, you can re-stimulate the cells with full medium w/ or w/o your cytokine treatment; If the cytokine really works on your cells, the treated cells should enter S phase and G2/M phase faster than the controls, and you can monitor this process by FACS during the next 48 h.
2. To determine if the cytokine affects the cell proliferation, you can also simply measure cell viability by either cell counting or MTT assays. You can also detect the changes of checkpoints genes, such as p53, p21, etc.
Hope it can help you!
Hi Giovanni,
Based on your data, I think there were be two possibility. One is that the cytokine slightly promoted the cell proliferation, and the addition experiment should be done to prove it. The other was the cytokine cause the production of polyploid cells, which may be found in the cell cycle distribution from the FCSC flow assay. I hope it will help you. Good luck.
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