Establishment of microglial cell cultures derived from postmortem human adult brain tissue: immunophenotypical and functional characterization.
Cell cultures have become an integral part of the daily routine in most biological research laboratories. Because they are very dynamic and highly accessible, cell cultures permit direct experimental manipulations where cause-effect relations can be more definitely assayed. We have developed cultures of microglial cells from rapid autopsies (range 3-10 hours) of nondemented elderly patients and Alzheimer's disease patients. Cultures were derived from the subcortical white matter, corpus callosum, and frontal, temporal, and occipital cortex. The adherent microglial cells were immunoreactive for CD68, CD45, CD11c, and major histocompatibility complex (MHC) class II markers, and were not immunoreactive for astrocyte or oligodendrocyte markers. In addition, some functional characteristics of the isolated microglial cells were also studied. Upon stimulation with lipopolysaccharide (LPS), microglial cells secreted pro- and antiinflammatory mediators, i.e., interleukin- (IL)-6, prostaglandin E2 (PGE2), and IL-10, indicating the functional capacity of cultured microglia.
de Groot
C J
CJ
Research Institute Neurosciences Vrije Universiteit Amsterdam, Department of Pathology, University Hospital, Vrije Universiteit, The Netherlands. cja.degroot@azvu.nl
Hulshof
S
S
Hoozemans
J J
JJ
Veerhuis
R
R
D000368:Aged; D000369:Aged, 80 and over; D000544:Alzheimer Disease / Q000473:pathology; D000911:Antibodies, Monoclonal; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002469:Cell Separation; D002478:Cells, Cultured; D016207:Cytokines / Q000096:biosynthesis; D015232:Dinoprostone / Q000096:biosynthesis; D005625:Frontal Lobe / Q000166:cytology; D000949:Histocompatibility Antigens Class II / Q000032:analysis / Q000276:immunology; D006801:Humans; D007150:Immunohistochemistry; D016130:Immunophenotyping; D017628:Microglia / Q000166:cytology* / Q000276:immunology; D008875:Middle Aged; D009836:Oligodendroglia / Q000166:cytology / Q000276:immunology
D000911:Antibodies, Monoclonal; D016207:Cytokines; D000949:Histocompatibility Antigens Class II; D015232:Dinoprostone