A murine model of xenotransplantation of human glioblastoma with imunosupression by orogastric cyclosporin

Abstract

Several animal experimental models have been used in the study of malignant gliomas. The objective of the study was to test the efficacy of a simple, reproducible and low cost animal model, using human cells of glioblastoma multiforme (GBM) xenotransplantated in subcutaneous tissue of Wistar rats, immunosuppressed with cyclosporin given by orogastric administration, controlled by nonimunosuppressed rats. The animals were sacrificed at weekly intervals and we have observed gradual growth of tumor in the immunosuppressed group. The average tumor volume throughout the experiment was 4.38 cm 3 in the immunosuppressed group, and 0.27 cm 3 in the control one (p<0.001). Tumors showed histopathological hallmarks of GBM and retained its glial identity verified by GFAP and vimentin immunoreaction. Immunosuppression of rats with cyclosporin was efficient in allowing the development of human glioblastoma cells in subcutaneous tissues. The model has demonstrated the maintenance of most of the histopathological characteristics of human glioblastoma in an heterotopic site and might by considered in research of molecular and proliferative pathways of malignant gliomas. Key words: animal model, Wistar rats, cyclosporin, glioblastoma, xenotransplant.Modelo murino de xenotransplante de glioblastoma humano com imunossupressão utilizando ciclosporina orogástrica RESUMO Vários modelos animais têm sido avaliados no estudo dos gliomas e até o momento nenhum pôde ser considerado ideal. O objetivo deste trabalho é verificar a eficácia de um modelo animal simples, reprodutível e de baixo custo. Utilizamos células humanas de glioblastoma multiforme (GBM) xenotransplantadas em ratos Wistar, submetidos a imunossupressão com ciclosporina administrada por via orogástrica. Células tumorais foram implantadas no tecido subcutâneo dos ratos imunossuprimidos com ciclosporina, sendo o controle feito em ratos não imunossuprimidos. Os animais foram sacrificados em intervalos semanais e foi observado crescimento progressivo do tumor no grupo imunossuprimido. A média do volume tumoral em todo o experimento foi de 4,38 cm 3 no grupo imunossuprimido e 0,27 cm 3 no grupo controle (p<0,001). Os tumores apresentavam características histopatológica do GBM e mantinham sua identidade glial, verificadas por imunoreação para GFAP e vimentina. A imunossupressão dos ratos com ciclosporina

Morphology of Human Glioblastoma Model Cultured in Ovo

“…Due to limited response of cultured tumour cells, a high demand for animal model studies is recognised. Most of the investigations using animal models are conducted on rodents (5). We propose an innovative model of the chicken embryo in ovo.…”

Tumour induction in BALB/c mice for imaging studies: An improved protocol

“…Iranpour et al suppressed the immune system of male C57BL/6 mice by combination of CsA, itraconazole and co‐amoxiclav 21 . Cunha et al reported xenotransplantation (cross‐species transplantation) of human cells of glioblastoma in rats by CsA treatment (5 mg/kg, daily) until the final step of experiments 8 . Du et al used CsA (10 mg/kg, daily) to develop xenografts of human hepatocarcinoma cells in C57BL/6 mice 22 .…”

“… 21 Cunha et al reported xenotransplantation (cross‐species transplantation) of human cells of glioblastoma in rats by CsA treatment (5 mg/kg, daily) until the final step of experiments. 8 Du et al used CsA (10 mg/kg, daily) to develop xenografts of human hepatocarcinoma cells in C57BL/6 mice. 22 According to the report of Rose et al, CsA (2–3 mg/kg) in combination with ketoconazole (10 mg/kg) was effective for the successful transplantation of xenologous skin in an immunosuppressed sheep model.…”

“…Since 1978, CsA has been frequently used both clinically and experimentally 4 . Many clinical researchers reported tumour xenograft development by modifying the CsA dose, administration route, combination with another drug and duration of treatment regimens 5–9 . CsA is metabolized in the liver by cytochrome P450.…”

“… 4 Many clinical researchers reported tumour xenograft development by modifying the CsA dose, administration route, combination with another drug and duration of treatment regimens. 5 , 6 , 7 , 8 , 9 CsA is metabolized in the liver by cytochrome P450. CsA metabolism can be decelerated by cytochrome inhibitors like diltiazem or ketoconazole.…”