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目的 研究C5 0株抗CEA杂交瘤细胞的抗体生成动力学 ,确定优化体外培养条件。方法 体外静态培养C5 0细胞 ,采用ELISA和细胞计数的方法检测培养上清抗体浓度和细胞数量 ,分析细胞生长与抗体分泌的关联性。采用体内复壮和添加新型免疫增强剂CpGODN的方法 ,尝试恢复长期体外培养的C5 0细胞的抗体生成能力。结果 C5 0细胞的抗体生成动力学为非生长关联型 ,经过体内复壮 ,C5 0细胞体外培养上清中抗体浓度显著提高 ,但活细胞密度或细胞活力均未发生明显变化。在培养基中添加一定浓度的CpGODN ,可以恢复长期体外培养的C5 0细胞的抗体生成能力。结论 根据细胞的抗体生成动力学特征 ,通过调节细胞的生长状态 ,可提高上清抗体浓度。根据抗体基因的组织特异性表达的机制 ,通过优化杂交瘤细胞的体外培养条件 ,能保持细胞正常的抗体生成能力 ,维持细胞高且稳定的上清抗体浓度
OBJECTIVE: To study the antibody production kinetics of C5 0 anti-CEA hybridoma cells and to optimize the culture conditions in vitro. Methods Cultured C5 0 cells were cultured in vitro. The antibody concentration and number of cells were detected by ELISA and cell counting. The correlation between cell growth and antibody secretion was analyzed. In vivo rejuvenation and adding a new immunostimulant CpGODN method, trying to restore the long-term in vitro C 50 cells antibody production capacity. Results The antibody production kinetics of C5 0 cells was non-growth-related. After in vivo rejuvenation, the concentration of antibody in the supernatant of C5 0 cells was significantly increased, but there was no significant change in viable cell density or cell viability. Adding a certain concentration of CpG ODN to the medium can restore the capacity of long-term culture of C5 0 cells in vitro. Conclusion According to the kinetics of cell antibody production, the concentration of antibody in the supernatant can be increased by adjusting the cell growth status. According to the mechanism of tissue-specific expression of antibody genes, by optimizing the in vitro culture conditions of hybridoma cells, the normal antibody production ability of cells can be maintained and the cell high and stable supernatant antibody concentration can be maintained