I would like to know if the DNA of a feeder cell layer, which was inactivated by mitomycin C, is amplified. I want to run a PCR on the cells cultured on the feeder layer and I'm afraid that the feeder cells will bias the outcome of the PCR.
On the other hand mitomycin C hinders cell proliferation by preventing replication of the DNA.
Ideally, you should pick individual cell colony grown on the feeder layer. This will ensure you have a purer cell population of interest when performing PCR and minimize feeder cell contribution
Action mechanisms of the MMC-treated feeder layer may vary among types of cells: for instance, a feeder layer of human foreskin fibroblasts but not that of mouse embryonic fibroblasts secretes basic fibroblast growth factor.
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4888025/
The DNA from feeder cells biases the outcome of PCR. If this is for stem cell cultures, colonies will be picked up with a minimal contamination of feeder cells. If stem cells are originated from human and feeder cells from mouse, contamination can be quantitated e.g., by a real time PCR for mouse mitochondrial DNA. Other approach may be replating of cells without a feeder layer prior to collection of cells.
A feeder layer (e.g., of cells inactivated by MMC or high dose ionizing irradiation) is useful to improve cell proliferation and plating efficiency. However, it is worthwhile keeping in mind the possibility that the results to be obtained reflect the bystander effects. For example, bystander effects of ionizing radiation can be broadly described as the occurrence of biological effects in cells that have not themselves been irradiated but have received signals from irradiated cells. Irradiated cells secrete a variety of soluble factors that include not only pro-mitogenic factors but also factors that may be toxic to cells of interest, which can affect other cells.
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