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

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.