细胞培养过程中支原体污染的防治与检测方法  

Treatment and detection of mycoplasma contamination during cell culture

作　　者：邢龙彬 刘长政[1] 焦晓磊[2] 刘彤[2] 杜智[2] 高英堂[2] 

机构地区：[1]天津医科大学第三中心临床学院,天津市300070 [2]天津市人工细胞重点实验室天津市肝胆疾病研究所天津市第三中心医院,天津市300170

出　　处：《世界华人消化杂志》2016年第10期1557-1564,共8页World Chinese Journal of Digestology

基　　金：天津市卫计委重点攻关基金资助项目,No.11KG112;天津市应用基础与前沿技术研究计划青年基金资助项目,No.15JCQNJC115000~~

摘　　要：目的:为高效便捷地去除细胞培养过程中支原体污染,采用多种抗生素和检测方法对污染细胞进行处理和检测.方法:分别采用Plasmocin、BM-Cyclin、MRA3种药物对污染细胞进行处理,以CLARK的一步法试剂、Biotool的快速检测试剂和自行研制的聚合酶链式反应(polymerase chain reaction,PCR)等3种方法进行检测.结果:一步法试剂检测显示Plasmocin处理14d后支原体消除,快速检测试剂检测显示BM-Cyclin处理21d彻底消灭支原体,而MRA经PCR检测发现彻底消除支原体需要14d;另外,3种药物交替使用清除支原体的效果更加显著.结论:通过使用3种抗生素和3种支原体检测方法,使细胞培养过程中的支原体污染得到有效控制,为更好地开展细胞研究奠定基础.AIM： To eliminate and detect the contamination of mycoplasma during cell culture effectively.METHODS： Three drugs（plasmocin, BM-Cyclin,and MRA） were used to treat the contaminated cells. CLRAK One Step Test Kit, Biotool Detection Kit and polymerase chain reaction（PCR） were used to detect the degree of contamination. RESULTS： Plasmocin, BM-Cyclin, and MRA could eliminate the contamination effectively.Assay with the CLARK Kit showed that mycoplasma disappeared completely on 14th day after treatment with plasmocin, the Biotool Detection Kit showed that the contamination disappeared completely on the 21st day after treatment with BM-Cyclin, and PCR showed that mycoplasma disappeared completely on the 14th day after treatment with MRA. The combination of the three drugs was more effective in the eradication of mycoplasma contamination.CONCLUSION： The combination of three antibiotics and three mycoplasma detection methods can effectively control mycoplasma contamination during cell culture.

关 键 词：支原体 细胞培养 防治 检测 

分 类 号：Q813.11[生物学—生物工程]