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Small‐scale isolation and characterisation of human peripheral blood monocytes
Author(s) -
Wynne Katherine M.,
Moore M.
Publication year - 1982
Publication title -
the journal of pathology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.964
H-Index - 184
eISSN - 1096-9896
pISSN - 0022-3417
DOI - 10.1002/path.1711380102
Subject(s) - ficoll , centrifugation , peripheral blood , differential centrifugation , stain , receptor , chemistry , endocytic cycle , isolation (microbiology) , substrate (aquarium) , yeast , biology , microbiology and biotechnology , immunology , biochemistry , staining , peripheral blood mononuclear cell , in vitro , endocytosis , ecology , genetics
A technique is described for the isolation and characterisation of monocytes from 15 ml of human peripheral blood. After density gradient centrifugation over Ficoll‐Hypaque, monocytes are purified further by substrate adherence under carefully defined conditions. The use of a simple microwell slide system permits the production of multiple small‐scale monolayer populations which can be characterised further in terms of their histochemical reactivity (combined stain for chloroacetate and non‐specific esterases); endocytic capacity (latex particles); Fc (sensitised ox erythrocytes) and C3 (serum‐coated yeast) receptor expression; and precursor maturation potential (7‐day cultures). Evidence of considerable cellular heterogeneity is presented.