Objective: To investigate the immunotherapy efficacy of fusion cells (dendritic-C6anti-TGF-β1 cells) in the treatment of intraeranial gliomas. Methods: Dendritic cells were isolated from rat bone-marrow precursor...Objective: To investigate the immunotherapy efficacy of fusion cells (dendritic-C6anti-TGF-β1 cells) in the treatment of intraeranial gliomas. Methods: Dendritic cells were isolated from rat bone-marrow precursors stimulated in vitro with granuloeyte-macrophage colony-stimulating factor (GM-CSF) and Interleukin-4 (IL-4). C6anti-TGF-β1 cells originally from C6 cell line of a rat glioblastoma were transfected with plasmid of TGF-β1 anti-sense gene. Fusions of dendritic cells and C6anti-TGF-β1 cells were prepared by polyethylene glycol (PEG). The DC/C6anti-TGF-β1 fusion cells were observed and confirmed by fight microscopy and scanning electron microscopy. Experimental rats were divided into three groups at random: C6 cells (Ⅰ), dendritic-C6anti-TGF-β1 fusion cells and C6 cells (Ⅱ) and IMDM medium only (Ⅲ). The cells were injected into right parietal lobe region of the rat with stereotaxic technique. Histology, tumor necrosis and survival time were evaluated. Results: Compared with the rats that received C6 cells (survival median time was less than 20 days, tumor region was seen in all fields of observed), the rats injected with dendritic-C6anti-TGF-β1 fusion cells and C6 cells got a more prolonged life span (more than 59 days), as well as less tumor region (5.01%-6.2%). There was no tumor necrosis, but some glias were seen in surroundings. All rats were survived and no necrosis was observed in negative control group. Statistical analysis showed that group Ⅱ had significant difference compared with group Ⅰ. Conclusions: Dendritic-C6anti-TGF-β1 fusion cells could prolong the life span of rats, providing a strategy to achieve an antitumor response against tumors in the central nervous system.展开更多
Objective: To establish a stable C6/EGFP glioma cell transfected with the human immunodeficiency virus line for studies on glioma. Methods: The C6 glioma cell line was type I (HIV-1) based lentivirus vector contai...Objective: To establish a stable C6/EGFP glioma cell transfected with the human immunodeficiency virus line for studies on glioma. Methods: The C6 glioma cell line was type I (HIV-1) based lentivirus vector containing two enhancer-promoters CMV and EF1α. Enhanced green fluorescent protein (EGFP)-positive C6 cells were sorted out by fluorescence-activated cell sort. Expression of EGFP was observed by fluorescent microscopy. EGFP gene in C6 genome was assessed by Polymerase chain reaction (PCR) and DNA sequencing. Original and transfected cells were compared biologically and cytomorphologically. Results: Lentivirus vector transfection produced up to 40% EGFP-positive cells. After fluorescence-activated cell sort selection, a pure cell line C6/EGFP was established. PCR and DNA sequencing revealed integration of EGFP gene in C6 cell genome. Analysis of cell characteristics revealed no difference between transfected and original cells. Conclusion: A C6/EGFP cell line expressing EGFP as a marker is established, in which the EGFP gene is integrated into the genome. This cell line can be served as a promising tool for further basic research and gene therapy studies.展开更多
文摘Objective: To investigate the immunotherapy efficacy of fusion cells (dendritic-C6anti-TGF-β1 cells) in the treatment of intraeranial gliomas. Methods: Dendritic cells were isolated from rat bone-marrow precursors stimulated in vitro with granuloeyte-macrophage colony-stimulating factor (GM-CSF) and Interleukin-4 (IL-4). C6anti-TGF-β1 cells originally from C6 cell line of a rat glioblastoma were transfected with plasmid of TGF-β1 anti-sense gene. Fusions of dendritic cells and C6anti-TGF-β1 cells were prepared by polyethylene glycol (PEG). The DC/C6anti-TGF-β1 fusion cells were observed and confirmed by fight microscopy and scanning electron microscopy. Experimental rats were divided into three groups at random: C6 cells (Ⅰ), dendritic-C6anti-TGF-β1 fusion cells and C6 cells (Ⅱ) and IMDM medium only (Ⅲ). The cells were injected into right parietal lobe region of the rat with stereotaxic technique. Histology, tumor necrosis and survival time were evaluated. Results: Compared with the rats that received C6 cells (survival median time was less than 20 days, tumor region was seen in all fields of observed), the rats injected with dendritic-C6anti-TGF-β1 fusion cells and C6 cells got a more prolonged life span (more than 59 days), as well as less tumor region (5.01%-6.2%). There was no tumor necrosis, but some glias were seen in surroundings. All rats were survived and no necrosis was observed in negative control group. Statistical analysis showed that group Ⅱ had significant difference compared with group Ⅰ. Conclusions: Dendritic-C6anti-TGF-β1 fusion cells could prolong the life span of rats, providing a strategy to achieve an antitumor response against tumors in the central nervous system.
基金supported by the National Natural Science Foundation of China(No.30640073)Beijing Municipal Natural Science Foundationand the Scientific Research Foundation for Returned Scholars,from Ministry of Education of china.
文摘Objective: To establish a stable C6/EGFP glioma cell transfected with the human immunodeficiency virus line for studies on glioma. Methods: The C6 glioma cell line was type I (HIV-1) based lentivirus vector containing two enhancer-promoters CMV and EF1α. Enhanced green fluorescent protein (EGFP)-positive C6 cells were sorted out by fluorescence-activated cell sort. Expression of EGFP was observed by fluorescent microscopy. EGFP gene in C6 genome was assessed by Polymerase chain reaction (PCR) and DNA sequencing. Original and transfected cells were compared biologically and cytomorphologically. Results: Lentivirus vector transfection produced up to 40% EGFP-positive cells. After fluorescence-activated cell sort selection, a pure cell line C6/EGFP was established. PCR and DNA sequencing revealed integration of EGFP gene in C6 cell genome. Analysis of cell characteristics revealed no difference between transfected and original cells. Conclusion: A C6/EGFP cell line expressing EGFP as a marker is established, in which the EGFP gene is integrated into the genome. This cell line can be served as a promising tool for further basic research and gene therapy studies.