线粒体DNA缺失细胞（ρ^0细胞）的构建与鉴定  

Construction and identification of mtDNA deletion cell line

作　　者：刘妍[1] 孔令平[1] 陈胜国[2] 汪华侨[2] 

机构地区：[1]广州医学院从化学院实验中心,广东从化510925 [2]中山大学中山医学院人体解剖学教研室脑研究室

出　　处：《解剖学研究》2010年第3期182-184,F0003,共4页Anatomy Research

基　　金：国家重点基础研究发展计划(973计划)(2006cb500700);广东省科技社会发展计划项目(2006B36004001;2006B36030003;2008B030301320)

摘　　要：目的探讨构建线粒体缺失细胞(ρ0细胞)的方法,为后继研究提供细胞模型。方法采用5μg/mL和7.5μg/mL两种浓度溴化乙锭持续作用于SH-SY5Y细胞构建线粒体缺失细胞,透射电镜观察和PCR法鉴定。结果 SHSY5Y细胞经过7.5μg/mL溴化乙锭持续作用95d后可见细胞线粒体肿胀变圆,基质消失,变成只剩外壳的"鬼影样"结构,线粒体嵴极少且变形;mtDNAPCR扩增未见目的条带。结论成功构建线粒体缺失细胞,为后继研究奠定细胞模型基础。Objective To explore the methods of construction of mtDNAdeletion cell line （ρ^0 cells） and provide the cell model in the followup study.Methods The mtDNAfree ρ^0 cell line used in this study was generated by incubating SHSY5Y cells in growth media containing 5 or 7.5 μg/ mL ethidium bromide （EtBr） for longterm incubation.Identification of ρ^0 cells were measured by electron microscopy and mtDNA PCR amplification.Results Complete transformation of SHSY5Y cells to mtD-NAdeletion ρ^0 cells required 95 days of incubation in 7.5 μg/ mL ethidium bromide.Mitochondria in ρ^0 cells became swollen and round, had grossly reduced and distorted cristae and essentially empty matrix giving it a ‘ghostlike＇ appearance.The target band was not seen after mtDNA PCR amplification.Conclusion Succeeding in construction of ρ^0 cells will provide cell model for the future indepth research.

关 键 词：ρ0细胞 溴化乙锭 线粒体 

分 类 号：R749.16[医药卫生—神经病学与精神病学]