I am trying to grow microglia culture for 10 days and after day 2/3 there are certain flat non-microglial contaminating cells observed. They are rapidly growing and some are even growing on top or in contact with microglia. I am trying to find a way to distinguish what these cells are and wanted to know if there are any ways of determining these cells? Could it potentially be fibroblasts or mesenchymal cells?
Hi Ankitha Johnas
Attaching images could have helped in identifying exactly the cells. However primary cells contamination includes,
- Connective tissue cells such as fibroblasts can arise from the meninges or blood vessels during tissue dissociation and can quickly overgrow the culture due to their rapid proliferation. Other than this Oligodendrocyte Precursor Cells (OPCs), immature cells that can differentiate into oligodendrocytes, which produce the myelin sheath that insulates neurons. These cells grow at a faster rate compared to primary microglia population.
Hope it helps,
Thanks,
To competently answer your question, one needs to know the method used for microglial purification. In majority of cases, the contaminating cells are astrocytes that are flat and proliferate at high rates. Astrocytes can be stained with antibodies recognizing glial fibrillary acidic protein GFAP (some of most reliable are from Millipore-Sigma (Cat. #G3893). Presence of fibroblast is also possible. Brain fibroblasts are trickier to stain because they do not have a universal marker.