Setting Up an In Vitro Model to Study Glial Response to Peripheric Immune Cells

MURTA V; RAMOS AJ

Cordoba

Congreso; Reunion Anual de la Sociedad Argentina de Investigacion en Neurociencias 2018; 2018

Sociedad Argentina de Investigación en Neurociencias

Healthy central nervous system provides limited interactionbetween parenchymal astrocytes and immune peripheralcells. After an ischemic event, the blood?brain barrier iscompromised and leukocytes are drawn to the lesion,where a complex immune response arises involving bothlocal and systemic cells. The detrimental or beneficial rolesof this recruitment are still discussed. The aim of the presentwork is to shed some light on the systemic cues associatedwith the commitment of astrocytes to specific activatingprofiles in response to peripheral immune cells. To undertakethis challenge, we set up an in vitro model where primaryrat glial cells are co-cultured with eGFPþ leukocytesisolated from adult Wistar-TgN(CAG-GFP)184ys rats. Fixedleukocytes were used to analyze the effect of surface molecules.Using immunofluorescence, we evaluated astrocytereactivity (GFAP), microglial activation (Iba1), and the formationof glial scar-like structures. Short-term (6 hr) andlong-term (72 hr) effects were studied. Astrocytes in contactwith both fresh and fixed leukocytes had a fibrillar morphologyand increased GFAP expression. Cellular retractionand reorganization was evident, and scar-like structureswere seen. Microglia in contact with leukocytes had an activated(round) morphology. These results indicate that bothsoluble factors and surface molecules in leukocytes are capableof inducing astrocytes? reactivity, but further research isnecessary to determine more specific pathways involved.This work was supported by PICT2015-1451-UBACYT.