Characterization of microglial cells and their response to stimulation in an organotypic retinal culture system

Citation
K. Mertsch et al., Characterization of microglial cells and their response to stimulation in an organotypic retinal culture system, J COMP NEUR, 431(2), 2001, pp. 217-227
Citations number
64
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Neurosciences & Behavoir
Journal title
JOURNAL OF COMPARATIVE NEUROLOGY
ISSN journal
0021-9967 → ACNP
Volume
431
Issue
2
Year of publication
2001
Pages
217 - 227
Database
ISI
SICI code
0021-9967(20010305)431:2<217:COMCAT>2.0.ZU;2-B
Abstract
An organotypic culture system of the early postnatal rat retina was develop ed to study microglial activation within a tissue environment. One day afte r tissue preparation, microglial cells of the ganglion cell/nerve fiber lay er revealed features of activation. Cells acquired an ameboid morphology as revealed by Bandeiraea simplicifolia lectin staining. Proliferation-as rev ealed by Ki67 immunocytochemistry-resulted in higher cell densities. In the supernatant, tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6) , and monocyte chemoattractant factor-1 (MCP-1) were detected by using spec ific enzyme-linked immunosorbent assay systems, activated microglia being t he most likely source of their release. After 6 days in vitro (div), microg lial cells regained their resting morphology, and cell counts returned to c ontrol levels. Concomitantly, the release activity decreased to undetectabl e levels. When slices were treated at this later stage of cultivation (>6 d iv) with bacterial lipopolysaccharide (LPS; 100 ng/ml for 24 hours), microg lial cells became activated, as revealed by a change in morphology. In para llel, the LPS treatment also resulted in high levels of TNF-alpha, IL-6, an d MCP-1 in the culture medium. Both the release from the tissue and the mor phological changes of the microglia were reversible. Seventy-two hours afte r LPS removal, only microglia with ramified morphology were found, and rele ase activities returned to baseline. These data suggest that the organotypi c culture of the retina is a useful model for studying microglial activatio n from its resting form. J. Comp. Neurol. 431:217-227, 2001. (C) 2001 Wiley -Liss. Inc.