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【目的】选择最佳灭菌方法,建立参环毛蚓细胞原代培养实验体系。【方法】通过4因素和3水平的正交试验法考察不同浓度比例抗生素组合液对参环毛蚓细胞的灭菌效果,比较不同抗生素种类和浓度对参环毛蚓细胞原代培养的影响。【结果】以抗生素组合液200 U/mL青霉素钠+400μg/mL硫酸链霉素+300 U/mL硫酸庆大霉素+5μg/mL两性霉素B对组织块的灭菌效果最佳,10 d污染率为0%,且细胞能保持较好活性,并贴壁生长。【结论】组合式灭菌法是参环毛蚓细胞原代培养的最佳有效灭菌方法,且操作简捷易行。
【Objective】 To select the best sterilization method and establish the primary culture system of Ranuncula pilosa. 【Methods】 The four-factor and three-level orthogonal test were used to investigate the effect of different concentrations of antibiotic combinations on the sterilizing of Cyclocarya palustris cells. The effects of different types and concentrations of antibiotics on the primary culture of Pheretima aspergillum were compared. 【Results】 The antibiotic combination solution 200 U/mL penicillin sodium + 400 μg/mL streptomycin sulfate + 300 U/mL gentamicin sulfate + 5 μg/mL amphotericin B had the best sterilization effect on tissue blocks, 10 The contamination rate of d is 0%, and cells can maintain good activity and adherent growth. [Conclusion] The combined sterilization method is the best effective sterilization method for the primary culture of P. cyclogoni cells, and the operation is simple and easy.