Cell cycle analysis

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gil
gil's picture
Cell cycle analysis

Hi everyone, I'm working on Glioblastoma multiforme (one of the most agressive brain cancer) with 20 microgram/ml THC induction. When I analyzed my 2 cell lines using the cell cycle analysis with several concentration of THC, i discovered that there is a G1 arrest when i use 10 microgram/ml THC at one cell line and 20 microgram/ml THC at the other. meaning-less to say that when i checked several concentrations at the MTT assay, 10 microgram/ml was elevated and did not show any decrease at the cell viability.
Does anyone know why? How can I explain this?

samm
samm's picture
This is probably simply a

This is probably simply a function of the assay you perform. The cell cycle assay with propidium iodide gives a snapshot of cell viability, death and cell cycle progression (which is why kinetic expts are very important in this type of assay) for individual cells in a population (if you perform a flow cytometric analysis). MTT on the other hand is merely a metabolic assay, which only indicates extent of viable cells in a bulk population.
G0/G1 arrest is NOT indicative of cell death/apoptosis - they can remain metabolically active and viable for quite some time. However, preventing cell division in any cancerous cell is half the job done!

gil
gil's picture
Thank you some much!

Thank you some much!

montgomj
montgomj's picture
Another assay you may want to

Another assay you may want to try is CFSE staining. You label your cells with this compound and then assay them at various time points by FACS. The fluorescence is diluted with each cell division giving a peak for each mitosis. This could help confirm your cell cycle arrest. You can find this on Invitrogen's website.