摘要
目的：通过对多种支原体污染去除方法治疗效果进行比较,找到适于人肝癌细胞系C3A去除支原体污染的最佳处理方法。方法通过聚合酶链反应（ PCR）检测C3A细胞培养物中支原体污染,并采用环丙沙星、加温处理、商品化支原体清除剂Plasmocure、人原代巨噬细胞体外共培养等方法对支原体污染样本进行处理,透射电镜（ TEM）和实时荧光定量PCR（ Q-PCR）对Plasmocure处理前后的细胞形态和基因表达进行比较分析。结果 Plasmocure成功清除支原体且无复发,环丙沙星清除支原体后复发,加温处理、巨噬细胞共培养法导致细胞死亡清除失败；电镜观察Plasmocure处理后细胞无支原体,生长良好,ALB、TF、CYP3A4表达较污染组明显增高（P〈0.01）。结论 Plas-mocure是支原体污染的有效处理方法并且能显著恢复细胞形态及基因表达。
Objective To evaluate the effectiveness of several methods in eliminating Mycoplasma contamination in cell culture of human hepatoma-derived cell line C3A. Methods PCR was performed to detect the Mycoplasmas contamination in cell cultures. The contaminated samples were treated by ciprofloxa-cin, heating, Plasmocure or co-culturing with macrophages. Transmission electron microscope （ TEM） and Q-PCR were used to comparatively analyze the cell morphology and gene expression before and after Plas-mocure treatment. Results Plasmocure succeeded in eliminating Mycoplasma contamination, while cipro-floxacin showed temporary efficacy. Heating and co-culturing with macrophages failed to eliminate Mycoplas-ma contamination. No Mycoplasma contamination in the Plasmocure-treated group was observed under TEM and the expression of ALB, TF and CYP3A4 genes were higher than the genes expressed in the contaminated group （P〈0. 01）. Conclusion Plasmocure treatment was effective in eliminating Mycoplasma contamina-tion in cell culture. Moreover, the cell morphology and gene expression in Plasmocure-treated group were re-stored to normal.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2016年第10期766-770,共5页
Chinese Journal of Microbiology and Immunology
基金
国家高技术研究发展计划（863计划）（2012AA020505）
国家自然科学基金（81470875）
广东省省级科技计划项目（20148020227002
20158090903069）
广东省自然科学基金项目（2014A030312013）
关键词
支原体污染
清除方法
细胞培养
Mycoplasma contamination
Elimination method
Cell culture
