CHARACTERIZATION OF INFILTRATING MACROPHAGES IN ORGANOTYPIC 3D CULTURE MODEL OF GLIOBLASTOMA

IVANA GISELE ESTECHO; MARIA JOSE GATTAS; ONIAS CASTELLANOS COLLAZO; MARÍA AMPARO LAGO HUVELLE; TOMÁS LAPORTE; MARÍA INÉS DÍAZ BESSONE; ANDREA E. ERRASTI; EUGENIO ANTONIO CARRERA SILVA; MARINA SIMIAN

Mar del Plata

Congreso; Reunión Conjunta SAIC SAI SAFIS 2018; 2018

SAIC-SAI-SAFIS

Glioblastoma multiforme (GBM) is one of the most devastating cancertypes with very limited therapeutic options and tumor-associatedmacrophages have been described to play a critical role in its growthand progression.The aim of this study was to set up a 3-dimensional (3D) organotypichuman model of GBM to characterize macrophages infiltrationand to identify potential therapeutic targets. We have used U-87 MGhuman glioblastoma cells together with human primary monocytes,or the THP-1 monocyte cell line. We established that hanging-dropcultures originating from ~3000 GBM cells resulted in reproduciblespheroids. Once they were formed, the spheroids were transferredinto the wells and co-cultured with 40.000 primary human monocytesor THP-1 cells. Monocytes invaded the spheroids as established bytracking with a live-cell fluorescent dye. Moreover, flow cytometryanalysis of dissociated spheroids co-cultured with monocytes during7 days, clearly shows CD11b+ macrophages skewed to M2 phenotypewith high levels of CD14, CD206, MERTK and CD163 comparedto the mean intensity fluorescence of monocyte-derived macrophagesalone (p<0.01). M2 polarization was also observed whenprimary monocytes were exposed to U-87 MG conditioned media in2D culture. The development of the co-culture-based 3D model ofGBM will allow us to test new therapeutic strategies that specificallytarget M2 macrophages under physiologically relevant settings.