Image-Based Assessment of Natural Killer Cell Activity against Glioblastoma Stem Cells

Glioblastoma (GBM) poses a significant challenge in oncology and stands as the most aggressive form of brain cancer. A primary contributor to its relentless nature is the stem-like cancer cells, called glioblastoma stem cells (GSCs). GSCs have the capacity for self-renewal and tumorigenesis, leading to frequent GBM recurrences and complicating treatment modalities. While natural killer (NK) cells exhibit potential in targeting and eliminating stem-like cancer cells, their efficacy within the GBM microenvironment is limited due to constrained infiltration and function. To address this limitation, novel investigations focusing on boosting NK cell activity against GSCs are imperative. This study presents two streamlined image-based assays assessing NK cell migration and cytotoxicity towards GSCs. It details protocols and explores the strengths and limitations of these methods. These assays could aid in identifying novel targets to enhance NK cell activity towards GSCs, facilitating the development of NK cell-based immunotherapy for improved GBM treatment. ### Competing Interest Statement The authors have declared no competing interest. * GBM : glioblastoma GSCs : glioblastoma stem cells NK cells : natural killer cells MHC-I : major histocompatibility class I EGF : epidermal growth factor bFGF : basic fibroblast growth factor IL-2 : interleukin-2 FBS : fetal bovine serum DPBS : Dulbecco’s Phosphate-Buffered Saline DMEM : Dulbecco’s Modified Eagle Medium DMSO : dimethyl sulfoxide ATCC : American Type Culture Collection