An optimized Matrigel-assisted orthotopic GL261 glioma model improves tumor engraftment and reproducibility.

BACKGROUND

Reliable and reproducible orthotopic GL261 glioma models are essential for preclinical glioblastoma research, particularly for evaluating therapeutic strategies in immunocompetent mice. However, conventional intracranial implantation of GL261 cells frequently results in variable tumor engraftment and heterogeneous tumor growth, limiting experimental reproducibility and increasing variability in therapeutic studies.

NEW METHOD: We developed an optimized Matrigel-assisted orthotopic GL261 glioma model by incorporating Matrigel into the conventional intracranial implantation procedure. Tumor establishment and progression were evaluated using bioluminescence imaging and micro-computed tomography, followed by histopathological and immunohistochemical analyses.

RESULTS

The optimized Matrigel-assisted approach achieved highly consistent tumor engraftment and significantly improved the reproducibility of intracranial tumor growth compared with the conventional implantation method. Histopathological analyses demonstrated intracranial GL261 tumors with characteristic histopathological features. Immunohistochemical staining demonstrated infiltration of CD3⁺ T lymphocytes and Iba1⁺ microglia/macrophages within the tumor tissues.

COMPARISON WITH EXISTING METHOD: Compared with the conventional orthotopic implantation method, the optimized Matrigel-assisted model provides more reliable tumor establishment and reduced inter-animal variability while demonstrating immune cell infiltration within tumor tissues.

CONCLUSIONS

An optimized Matrigel-assisted orthotopic GL261 glioma model significantly improves tumor engraftment and experimental reproducibility in immunocompetent mice. This methodological improvement provides a practical and reproducible experimental platform for preclinical glioma studies requiring consistent intracranial tumor establishment.