植物血凝素-ECM830细胞融合法融合黑色素瘤细胞

Fusion of Melanoma Cells Using A Modified Phytohaemagglutinin-ECM830 Fusion Method

目的以黑色素瘤细胞为模型,研究高效率肿瘤细胞融合方法。方法慢病毒荧光标记黑色素瘤细胞,ECM830电融合法及改良的植物血凝素(PHA)-ECM830电融合仪细胞融合法融合黑色素瘤细胞,流式分选黑色素瘤融合细胞,碘化亚啶染色法测定DNA含量。结果荧光标记带有绿色和红色荧光的黑色素瘤细胞,通过PHAECM830细胞融合法获得黑色素瘤融合细胞,细胞融合效率较ECM830电融合法从0.50%提升到7.18%,流式分选得到单一的黑色素瘤融合细胞,DNA含量分析显示黑色素瘤融合细胞的DNA含量翻倍,体内外实验表明黑色素瘤融合细胞增殖速度降低。结论运用改良的PHA-ECM830细胞融合法成功高效得到黑色素瘤融合细胞,黑色素瘤融合细胞增殖速度降低。

Objective To study melanoma cell fusion and find a highly efficient fusion method for tumor cells. Methods Melanoma cells were labeled with green fluorescent protein and red fluorescent protein, respectively, and fused by a modified phytohaemagglutinin （PHA） -ECM830 fusion method. Melanoma fusion cells were selected by the fluorescence activated cell sorting. DNA content was determined by propidium iodide staining. Results Melanoma cells were labeled with green fluorescent protein and red fluorescent protein markers and successfully fused through PHA-ECM830 fusion method. The fusion efficiency （7. 18% ） was much higher compared with ECM830 electricfusion method （0. 50% ） . Melanoma fusion cells were successfully obtained by the fluorescence activated cell sorting. DNA content was doubled in melanoma fusion cells compared to B16-F10 melanoma cells. The proliferation rate of melanoma fusion cells was significantly decreased in vitro and in vivo. Con- clusions We successfully obtained the melanoma fusion cells by the improved PHA-ECM830 fusion meth- od. The proliferation rate of melanoma fusion cells dramatically decreases.