Establishment of an orthotopic xenograft model of glioma with dual fluorescent labels

  Objective: To establish a stable, real-time monitoring of orthotopic glioma tumor model in nude mice.

  Methods: U251 glioma cells were infected with lentivirus carrying luciferase-Luc and green fluorescent protein-GFP genes, and cell lines stably expressing GFP-Luc fluorescence were screened by flow cytometry , And evaluate whether the proliferation, migration and invasion ability of fluorescent cells has changed through CCK-8 experiment, cell cycle experiment, Transwell tumor migration and invasion experiment; inoculate the cells into the caudate nucleus of nude mice to establish orthotopic transplantation of glioma Tumor model, using a mouse live imaging system to monitor the growth of tumors in the brain, and evaluate the pathological characteristics and tumor-forming ability of cells in the nude mouse brain through paraffin sections and HE staining.

  Results: The U251 glioma cell line and animal model stably expressing GFP fluorescence and luciferase fluorescence were successfully constructed. The lentiviral integration did not change the cell proliferation, migration and invasion ability; the model growth cycle was moderate, the tumor formation rate was high, and the tumor was in place. The intracranial growth is stable, and the HE section is consistent with the characteristics of human glioma.

  Conclusion: Compared with traditional cells, dual-fluorescence-labeled glioma cells are more conducive to the experimental study of glioma animal models; U251-GFP-Luc glioma cell nude mouse model has the same tumor growth and pathological characteristics as human glioma. Tumors are similar, and tumor growth can be observed in real time, which can be used as an ideal animal model for glioma experimental research.