| Objective Isolating and Culturing glioma stem cells (GSCs) from C6glioma cell line and cultured them in vitro, Identified the stem cell related markers. Compare the tumorigenicity between C6glioma cell and glioma stem cells Isolated from C6glioma cell line. Extracting the monocytes cells from rat bone and were induced to differentiate into dendritic cells.Two kinds of Anti-cancer DC vaccines were prepared by co-incubating with the whole tumor antigen from C6glioma cell line or glioma stem cells; To established the rat brain glioma model in wistar rat use the C6glioma cell, Two kinds of Anti-cancer DC vaccines were injected through tatl vein, the living span of rats with brain glioma were observed and analysis.Methods Culturing C6glioma cell in vitro, C6cell in logarithmic phase was cultured in serum-free medium to obtain glioma stem cells. Proliferation, colony formation and differentiation were analyzed. Glioma stem cells and C6glioma cell were subcutaneous inoculation into correspond rat. Compare the tumorigenicity use the PET-CT. Extracting the monocytes cells from rat femurs and induced into dendritic cells used cytokines.Prepared GSCs and C6cells antigen by freezing-melt, Made Anti-cancer DC vaccines by co-incubating with the whole tumor antigen obtain from GSCs or C6cells. Establishment intracranial glioma model by stereotactic inoculation of C6glioma cell line in rats. The living and life span of rats with glioma were observed. The examinations such asMRI, HE staining were used to observe the growth of tumor. Anti-cancer DC vaccines were injected through tail vein, compared the survival time and analysis survival. Results Glioma stem cells can form tumor spheres in serum-free medium and suspension growth. The expression of stem cell markers, sush as CD133and Nestin, were detected by immunofluorescence.The cell have great self-renewal and proliferation capability. The tumorigenicity was stronger than C6glioma cell. The dendritic cells have typical long dentrite in morphology and highly expressed DC markes,such as OX62. Through stereotactic inoculation, the rate of tumor formation was very high. The growth characteristics and pathological features of tumor is same to human glioma. The existence periods of rats injected DC vaccines use GSCs are longer than rats injected other DC vaccines.Conclusion DC loaded with Glioma stem cells angtine can provide a higher level of immunity protect against gliomas than DC vaccines made by C6glioma cell angtine. |
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