I need to find a basic protocol for preliminary work to isolate monocytes or macrophages from whole blood.
Current Protocols in Immunology, UNIT 7.6A "Isolation of Human Monocyte Populations", DOI: 10.1002/0471142735.im0706as70
I used to draw up to 500 ml of blood from a colleague in the US who needed only lymphocytes for his Science and Nature papers. Another colleague of mine who collaborated with me in cell adhesion and chemokines somehow collected the monocytes by having those cells just settle for I think about 2 hours on cell culture flasks... Not sure if this could be helpful for you.
EDIT: the non-adhering cells were washed away, the adhering cells were used.
I know that this is the way to separate peritoneal macrophages from total peritoneal lavage by just let them adheare 1-2 h and wash away the rest (WARM media!).
Are you sure monocytes are similiar adherent?
COLD media for seeding (otherwise they will stick to the falcon tube), WARM media for washing in the flask upon adhearence..
I am sure we only used huge amounts of blood samples from healthy donors (i.e. lab members) for getting human lymphocytes. I once even sold my own blood for my colleagues final experiment and a figure in a Science paper. We then were happy to use "waste" to isolate the monocytes, but those similarly fascinating results were never published...
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