Characterization of glioma cell line characteristics via antibody labeling in an orthotopic zebrafish model
Loading...
Date
2016-05
Authors
Journal Title
Journal ISSN
Volume Title
Publisher
The Ohio State University
Abstract
Glioblastoma (GBM) is a highly aggressive form of brain cancer with limited efficacious treatments and thus a low survival time in patients. To further understand the biology of GBM tumors, we utilized immunohistochemistry in an orthotopic xenotransplantation model of GBM in zebrafish. Patient-derived GBM cells were cultured and transplanted into larval animals to recapitulate the disease. Two cell types, GBM9 neurospheres and adherent cells, were characterized using markers of proliferation, differentiation, and stemness. Adherent cells were found to express higher levels of proliferation compared to neurospheres. While adherent cells displayed high levels of vimentin and glial fibrillary acidic protein (GFAP) at all time points, neurospheres expressed low levels of these markers early on that increased significantly over time in vivo. Conversely, neurospheres expressed high levels of Sox-2 positive cells early on with these populations decreasing over time. Adherent cells displayed low levels of Sox-2 positive cells over time, consistent with the static nature seen in GFAP and vimentin expression and opposite to the dynamic nature of neurospheres. Simultaneous labeling of multiple markers was utilized to further understand the relationship between the division rates of different cell populations. A small population of differentiated vimentin positive neurospheres was found to be dividing at a 5 dpt, while a majority of sox-2 positive neurospheres were dividing at the same time point. The number of dividing stem cells increased over time. Early on, a majority of GFAP positive cells were found to be proliferating but this number decreased significantly by 10 dpt. The results provide a deeper understanding of the biology of GBM tumors, specifically in relation to characteristics displayed by patient tumors.
Description
Keywords
glioblastoma, zebrafish, neurosphere, glioma, immunohistochemistry