I am working with primary cell culture at the moment and we have had issues with RBC contamination for the purposes of visualization of our cultured cells. Additionally, I've noticed pH changes (more acidic) with prolonged RBC contamination, but I'm having difficulty finding published data on what exactly the detrimental effects of RBC contamination are on cell culture.
Any help/references would be great!
RBCs cannot not be included in any cell culture since they are metabolizing nutrients and creating extra waste,causing definite pH changes. If the cancer cells are not attachable and making cell clumps during culture, transfer into 100mm dish containing PBS. Whirl the dish gently 3~4 times under a stereo microscope. You will see the large clumps will be collected in the middle of the dish, the RBCs will disperse at the periphery of the dishe due to the weights. Recover the clumps via a flame-polished (narrow pore now) Pasteur pipette in the middle. Or you could lyse the RBCs after centrifugation if the cancer cells are single cell suspended. (you can find it in Google)
Good luck.
Thanks so much! The pH changes caused by RBC metabolism were what I figured (and showed via pH sensitive color change in the media). But I didn't know if there was anything else worth noting about how they could be affecting cancer cell growth in vitro.
Much appreciated!
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