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Macro and microglial retinal cells in rat organotypic cultures
Author(s) -
GALLEGO B,
ROJAS B,
SALAZAR JJ,
RAMíREZ AI,
DIERSTEIN M,
DE HOZ R,
TRIVIñO A,
RAMíREZ JM,
UEFFING M,
ARANGOGONZáLEZ B
Publication year - 2013
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2013.t009.x
Subject(s) - retinal , microglia , retina , immunostaining , astrocyte , glial fibrillary acidic protein , pathology , in vivo , biology , vimentin , microbiology and biotechnology , cell culture , neuroglia , immunohistochemistry , neuroscience , immunology , medicine , central nervous system , inflammation , biochemistry , genetics
Purpose Organotypic retinal cultures constitute a useful tool to perform preclinical drug testing. The aim of the present study was characterize the morphologic changes in macro‐ and microglial cells in this culture system. Methods Retinas were isolated from 7‐day‐old Crl:CD(SD) rats with the retinal pigment epithelium attached. Retinal explants were cultured for 4, 10, and 14 days (DIV4, DIV10, and DIV14). Cryosections and whole‐mounts of age‐matched control and cultured retinas were used to analyze macro‐ and microglial cells by immunostaining using antibodies against GFAP, vimentin and CD11‐b. Results In comparison with in vivo, in DIV4 and DIV10 cultures, GFAP‐positive astrocytes were more robust, the astrocytic network being thicker in some retinal areas while in other regions astrocytes were sparsely distributed. Few thin GFAP‐positive astrocytes were observed at DIV14. Müller cells in the cultures exhibited GFAP up‐regulation in comparison with in vivo retinas. CD11‐b+ microglial cells at DIV4 and DIV10 showed more robust somas and thicker and more retracted processes than in vivo. Overall, at DIV14 CD11‐b+, microglial cells exhibited a rounded morphology. Conclusion In rat organotypic culture both macro‐ and microglial cells showed progressive changes: i) reactive macrogliosis, ii) rearranged astrocytic distribution, iii) GFAP up‐regulation in Müller cells, and iv) microglial activation. Given that this glial response is a hallmark of several retinal diseases, organotypic retinal culture is a valuable resource for future investigations in retinal degenerative processes and therapy.