Hello, good morning.
I would like to know which plate I can use to infect fresh PBMC with virus and culture for 24 hours.
I have tried using 12-well and 24-well plates, but many cells adhere and others remain suspended even after centrifugation. Since the cells do not form pellets after centrifugation, I am losing many during washes after virus infection. I need to preserve all cells, especially NK cells.
Your question is confusing. PBMC are blood cells that have mononucleated cells separated from the PMNs. As such PBMC are basically 60 to 80% lymphocytes and, ~30% monocytes. The monocytes will stick to almost any surface. The lymphs are non-adherent. Monos, once they stick, require a half stick of dynamite to remove them. (humor) If you want to work with lymphs remove the monos by non-specific adherence and collect the non-adherent lymphs. If you want to focus on NK cells isolate them with magnetic beads. If you want to culture all mononuclear cells use Teflon bags and the monos will not stick.
This is a superficial answer. It can go much deeper and I am sure someone else will extend it. You can also follow with a question and I can clarify as needed.
Thank you very much for your response. I will try to explain my question better. Maybe I will post it again.
In my study, I intend to infect PBMCs with SARS-CoV-2. After that, I will analyze NK cells by flow cytometry to check if their phenotype changes or if they show degranulation.
Now, I found an alternative. After the infection, I need to perform washes. I will aspirate the entire supernatant from the plate and transfer it to a 1.5 mL tube, then centrifuge it. After that, I will discard the supernatant and resuspend the cell pellet before placing it back into the plate. 24 hours after infection, I will collect the total supernatant and analyze the NK cells by flow cytometry.
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