I am currently performing cell cycle analyses using flow cytometry and was wondering what a double G0/G1 peak might indicate. It has appeared rather consistently in my data and is usually accompanied by an accumulation of cells in S phase, though not always. It is not a sub G0/G1 peak so much as it is a "notch" or "hump" in the usual G0/G1 peak. In the corresponding contour plot it almost looks as if there are two populations of G0/G1 cells.
I have read that it could represent variations in ploidy but upon talking with a cell cycle expert, they didn't think that was the case. Are there any other circumstances a phenotype like this would occur?
I should add: I am using a well-established trophoblast cell line of which I have knocked down a gene of interest.
It's unlikely there are two different cell populations.
The key may be related to the cell line.
Trophoblast cell lines have become an invaluable tool to study various aspects of trophoblast biology. These cell lines are more resilient when compared to cultures of primary trophoblast cells, including extended lifespans in culture, and are thus advantageous for studies encompassing more detailed investigations of trophoblast function and molecular biology. Trophoblast cell lines can be established from primary cultures by spontaneous in vitro selection or by ectopic expression of oncoproteins that confer immortality. Choriocarcinoma cells are transformed populations of trophoblast cells that readily adapt to in vitro conditions. However, because of these traits, data obtained from these manipulated or transformed cell lines must be interpreted with caution. The trophoblast cell lines have been removed from their in vivo environment for many passages and some of their properties may be distorted.
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