Setting up an in vitro model to study glial response to peripheric immune cells

ALBERTO JAVIER RAMOS; VERONICA MURTA

Córdoba

Congreso; XXXIII Congreso Anual de la Sociedad Argentina de Investigación en Neurociencias; 2018

Sociedad Argentina de Investigación en Neurociencias

Healthy CNS provides limited interaction between parenchymal astrocytes and immune peripheral cells. After an ischemic event the BBB is compromised and leukocytes are drawn to the lesion, where a complex immune response arises involving both local and systemic cells. The detrimental or beneficial roles of this recruitment are still discussed. The aim of the present work is to shed some light on the systemic cues associated with the commitment of astrocytes to specific activating profiles in response to peripheral immune cells. To undertake this challenge, we set up an in vitro model were primary rat glial cells are co-cultured with eGFP+ leukocytes isolated from adult Wistar-TgN(CAG-GFP)184ys rats. Fixed leukocytes were used to analyze the effect of surface molecules. Using immunofluorescence, we evaluated astrocyte reactivity (GFAP), microglial activation (Iba1), and the formation of glial scar-like structures. Short term (6 hs) and long term (72 hs) effects were studied. Astrocytes in contact with both fresh and fixed leukocytes had a fibrillar morphology and increased GFAP expression. Cellular retraction and reorganization was evident, and scar-like structures were seen. Microglia in contact with leukocytes had an activated (round) morphology. These results indicate that both soluble factors and surface molecules in leukocytes are capable of inducing astrocytes? reactivity, but further research is necessary to determine more specific pathways involved. PICT2015-1451, UBACYT.