I have isolated cells from muscle tissue, culture them in adherent flasks and want characterize them. CD56 is a marker for myoblasts, but also for natural killer cells and macrophages. Is it expected to have contaminating population of NKC or macrophages present among myoblasts to test positive for CD56? The cells are characterized after the expansion on plastic.
I have never cultured myocytes, but I have worked with macrophages and NK cells. Macrophages and NK cells co-express CD45 [also known as hematopoietic cell phosphatase] and C56. You can exclude them by excluding CD45+ cells from your analysis. Of the two, I would expect macrophages to be more prevalent than NK cells.
That said, I would not expect NK cells to grow under the conditions that support myocytes. NK cells need the cytokine IL-12 to grow. Macrophages are more plastic in their growth requirements, but they do need a source of either GM-CSF or M-CSF, which I also suspect are not present in myocyte culture conditions.
Hi Maria Olenic , agreeing to the answers above; in this case only macrophages are the adherent cell type that could be active in initial part of your culture. But in absence of any cytokine stimulation they should not mature and subsequently die, as you passage your cells. Very simple way is to check this (before you use your cells) will be to run a quick flow cytometry test. Culture should be CD45- (for any immune cell contaminant) and CD14-/CD16- (for macrophages).
Good luck!
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