Characterization and comparison of human glioblastoma models

Abstract

Glioblastoma (GBM) is one of the deadliest cancers. Treatment options are limited, and median patient survival is only several months. Translation of new therapies is hindered by a lack of GBM models that fully recapitulate disease heterogeneity. Here, we characterize two human GBM models (U87-luc2, U251-RedFLuc). In vitro, both cell lines express similar levels of luciferase and show comparable sensitivity to temozolomide and lapatinib exposure. In vivo, however, the two GBM models recapitulate different aspects of the disease. U87-luc2 cells quickly grow into large, well-demarcated tumors; U251-RedFLuc cells form small, highly invasive tumors. Using a new method to assess GBM invasiveness based on detecting tumor-specific anti-luciferase staining in brain slices, we found that U251-RedFLuc cells are more invasive than U87-luc2 cells. Lastly, we determined expression levels of ABC transporters in both models. Our findings indicate that U87-luc2 and U251-RedFLuc GBM models recapitulate different aspects of GBM heterogeneity that need to be considered in preclinical research.

Treatment of Orthotopic U251 Human Glioblastoma Multiforme Tumors in NRG Mice by Convection-Enhanced Delivery of Gold Nanoparticles Labeled with the β-Particle-Emitting Radionuclide, 177Lu

“…Human GBM tumors were established by stereotaxic inoculation of luciferase-transfected U251-Luc human GBM cells into the right cerebral hemisphere of the brain in female 6–8 week-old NRG mice (Cancer Stem Cell Colony, Princess Margaret Cancer Centre) . U251-Luc tumors recapitulate the most important histopathologic features of human GBM, especially an infiltrative pattern of invasion and palisading necrosis . NRG mice were used to establish U251-Luc tumors and to study the effectiveness and normal tissue toxicity of 177 Lu-AuNPs because other immunocompromised strains of mice such as NOD SCID mice harbor the Prkdc Scid mutation in DNA repair that results in unusual sensitivity to radiation .…”

“… 26 U251-Luc tumors recapitulate the most important histopathologic features of human GBM, especially an infiltrative pattern of invasion and palisading necrosis. 27 NRG mice were used to establish U251-Luc tumors and to study the effectiveness and normal tissue toxicity of 177 Lu-AuNPs because other immunocompromised strains of mice such as NOD SCID mice harbor the Prkdc Scid mutation in DNA repair that results in unusual sensitivity to radiation. 28 Luciferase-transfected U251-Luc human GBM cells were provided by Dr. James Rutka (Hospital for Sick Children, Toronto, ON, Canada) and were previously inoculated into the brain in NOD scid gamma (NSG) mice to establish orthotopic human GBM tumors.…”

Photobac derived from bacteriochlorophyll-a shows potential for treating brain tumor in animal models by photodynamic therapy with desired pharmacokinetics and limited toxicity in rats and dogs

“…The in vitro PDT efficacy of Photobac was investigated in the three cell lines: U87 [ 21 ], MG-human epithelial glioblastoma [ 22 ], C6-rat glial cell glioma [ 23 ] and F98-rat glial cell undifferentiated malignant glioblastoma [ 24 ]. For in vivo studies, U87 cells were subcutaneously implanted in SCID mice and C6 tumor cells were implanted in the brain of rats.…”

Optimization, Characterization, and Comparison of Two Luciferase-Expressing Mouse Glioblastoma Models

“…Luciferase expression of GL261 Red-FLuc and TRP-mCF cells was verified using a modified protocol from that previously described [17]. Briefly, 5000, 10,000, 15,000, and 20,000 cells/well were seeded in black clear-bottom 96-well plates (Corning, Corning, NY, USA) and incubated overnight (37 • C, 5% CO 2 ).…”

“…Luciferase expression allows for in vivo bioluminescent imaging of tumors in live animals, which yields real-time quantitation of the relative tumor size and treatment response while reducing animal usage and increasing throughput [15]. The most common luciferase reporters include Luc (original), Luc2 (next-generation codon-modified), and Red-FLuc (latest generation red-shifted, highest intensity, and suitable for deep tissue imaging) [16,17]. However, cancer cells expressing reporter proteins, including luciferase, can be immunogenic, which is associated with spontaneous tumor regression [18,19].…”