Single-cell gene expression of a glioma cell line (GL261) cultured adherently or as neurospheres

GL261 cells are a widely-used model system for glioma tumors and can be cultured adherently or as free-floating cell aggregates known as neurospheres. Neurospheres can be implanted into mice where they readily form tumors. This suggests that neurospheres may more closely resemble the in-vivo tumor phenotype. Our study has two primary objectives: 1) To determine differences in gene expression between adherent cells and the tumor-forming neurospheres and 2) To determine the extent of homogeneity in these cell populations. This project was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health (Grant #P20GM103408) along with a Murdock College Science Research Program Grant (SR-202119904) from the M.J. Murdock Charitable Trust. Overall design: GL261 cells were cultured adherently in DMEM with 10% FBS and 1% penicillin-streptomycin. Adherent cells were trypsinzed, collected from culture flasks, and analyzed using scRNA-Seq. Neurosphere formation was induced with the addition of epidermal growth factor (EGF, 20 ng / ml), fibroblast growth factor (FGF, 20 ng / ml) and B-27 (1:50) to the culture media. Neurospheres were collected after 12 days, trypsinized, and analyzed using scRNA-Seq.