Establishment of a novel neuroglioma transplantation model in rats

Objective:To establish a novel neuroglioma transplantation model in rats to demonstrate the dynamic growth of tumor.Methods:pEGFP-N1,a eukaryotic plasmid containing enhanced green fluorescent protein(EGFP) gene,was transfected into the rats glioma cell line C6 with the help of liposome lipofectamine.Then the cell clones(C6-gfp) stably expressing EGFP was obtained through neomycin G418 screening and subcloning.The growth of C6 and C6-gfp cells were observed using Cell Counting Kit-8(CCK-8).Then the C6-gfp cells were transplanted into Wistar rat's cranium.The tumor formation was observed using B ultrasound and H-E staining.The EGFP expression in the tumor was detected using fluorescence stereomicroscope and anti-GFP immunohistochemistry.Results:Flow cytometry showed that 97.7% of the screened C6-gfp cells were positive of EGFP.There was no obvious difference in the growth curves of C6 and C6-gfp cells(P0.05).The tumor formation rate in the rat's cranium was 70% after C6-gfp transplantation.The dynamic growth of tumor could be observed using B ultrasound and the expression of green fluorescence could be observed under the fluorescence stereomicroscope.H-E staining showed large nuclei in glioma cells,deep dying with more nuclear divisions.New vessels could be seen in the tumor.Anti-GFP immunohistochemistry showed GFP-positive cells with distinct intensity.Conclusion:A C6-gfp cell line stably expressing GFP has been successfully established in the present study and it can readily form tumors in the cranium,which lays a foundation for further study of prevention and treatment of glioma.