Glioblastoma and cerebral organoids: development and analysis of an in vitro model for glioblastoma migration. Issue 4 (18th February 2023) - Fedorova, Veronika
Abstract : It is currently challenging to adequately model the growth and migration of glioblastoma using two‐dimensional (2D) in vitro culture systems as they quickly lose the original, patient‐specific identity and heterogeneity. However, with the advent of three‐dimensional (3D) cell cultures and human‐induced pluripotent stem cell (iPSC)‐derived cerebral organoids (COs), studies demonstrate that the glioblastoma‐CO (GLICO) coculture model helps to preserve the phenotype of the patient‐specific tissue. Here, we aimed to set up such a model using mature COs and develop a pipeline for subsequent analysis of cocultured glioblastoma. Our data demonstrate that the growth and migration of the glioblastoma cell line within the mature COs are significantly increased in the presence of extracellular matrix proteins, shortening the time needed for glioblastoma to initiate migration. We also describe in detail the method for the visualization and quantification of these migrating cells within the GLICO model. Lastly, we show that this coculture model (and the human brain‐like microenvironment) can significantly transform the gene expression profile of the established U87 glioblastoma cell line into proneural and classical glioblastoma cell types. Abstract : Modeling glioblastoma in vitro remains a major challenge. This study established a protocol for coculturing cerebral organoids and glioblastoma spheroids that allows studying the glioblastoma under conditions resembling in vivo state. Approaches for visualization and evaluation of glioblastoma cell migration were introduced. Importantly, mRNA‐seq showed that glioblastoma cells cocultured with cerebral organoids changed their gene expression toward neural cell identity.