In my cell culture i have GFP positive and negative cells. I would like to know if there is a difference in cell proliferation rate in positive compared to negative cells without sorting the cells.
Normally we just count the cells to find out the proliferation rate but i have a mixture of positive and negative cells. I would like to compare the two. Could i use FACS somehow to gate the positive cells and find out if the cell went through more dividing cycles as the negative control?
Any suggestions?
I have found 3H-thymidine, but after reading a paper it seems quite unreliable.
You can use Edu incorporation and staining. You will get 4 populations all cells that are divided after the addition of Edu would be GFP/Edu positive or for non-GFP population GFP negative/Edu positive and cells that have not divided GFP-positive Edu negative and GFP negative-edu positive.
best way to check the difference in their cell proliferation status is to sort out the GFP positive cells in FACS and culture these cells, also, rest of the negative GFP population culture separately and grow them for couple of passage to check that cell are not loosing GFP population and preform the cell proliferation assay in both the cells.
As mentioned above, i can't sort the cells. It has to be done with the mixture of cells.