R. Timothy Bentley

High‐grade glioma is an aggressive cancer that occurs naturally in pet dogs. Canine high‐grade glioma (cHGG) is treated with radiation, chemotherapy or surgery, but has no curative treatment. Within the past eight years, there have been advances in our imaging and histopathology standards as well as genetic charactereization of cHGG. However, there are only three cHGG cell lines publicly available, all of which were derived from astrocytoma and established using methods involving expansion of tumour cells in vitro on plastic dishes. In order to provide more clinically relevant cell lines for studying cHGG in vitro, the goal of this study was to establish cHGG patient‐derived lines, whereby cancer cells are expanded in vivo by injecting cells into immunocompromized laboratory mice. The cells are then harvested from mice and used for in vitro studies. This method is the standard in the human field and has been shown to minimize the acquisition of genetic alterations and gene expression changes from the original tumour. Through a multi‐institutional collaboration, we describe our methods for establishing two novel cHGG patient‐derived lines, Boo‐HA and Mo‐HO, from a high‐grade astrocytoma and a high‐grade oligodendroglioma, respectively. We compare our novel lines to G06‐A, J3T‐Bg, and SDT‐3G (traditional cHGG cell lines) in terms of proliferation and sensitivity to radiation. We also perform whole genome sequencing and identify an NF1 truncating mutation in Mo‐HO. We report the characterization and availability of these novel patient‐derived lines for use by the veterinary community.