title: Cellular models for mitochondrial DNA-based diseases: lymphoblastoid cell lines and transmitochondrial cybrids.
abstract: Mitochondrial DNA (mtDNA) mutations cause a variety of mitochondrial DNA-based diseases which have been studied using Lymphoblastoid cell lines (LCLs) and transmitochondrial cybrids. Individual genetic information is preserved permanently in LCLs while the development of transmitochondrial cybrids provide ex-vivo cellular platform to study molecular mechanism of mitochondrial DNA-based diseases. The cytoplasmic donor cells for previous transmitochondrial cybrids come from patient's tissue or platelet directly. Here, we depicted in details the principle, methods and techniques to establish LCLs from frozen peripheral bloods harboring mitochondrial 4401G > A mutation by infection of Epstein Barr virus, and then to generate cybrids using ρ0206 and LCLs. The process of establishing these two cellular models was summarized into four steps as follows: (1) Generation of LCLs; (2) Transformation; (3) Selection; (4) Verification. To faithfully represent the function of mtDNA mutation, we analyzed and identified the sites of mtDNA mutations and copy numbers of each cellular models as well as the karyotype of transmitochondrial cybrids. Those clones with consistent parameters were selected for preservation and future analysis of the function of point mutations of mtDNA. Although these two cellular models play important roles in understanding molecular mechanism of mitochondrial DNA-based diseases on the cellular level, their limitations should be considered when elucidating the character of tissue specificity of mitochondrial DNA-based diseases.
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Ji-ji
authors.initials: JJ
authors.lastname: Sun
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Xiao-xu
authors.initials: XX
authors.lastname: Zhao
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Li-hua
authors.initials: LH
authors.lastname: Qiao
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Shuang
authors.initials: S
authors.lastname: Mei
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Zhi-peng
authors.initials: ZP
authors.lastname: Nie
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Qing-hai
authors.initials: QH
authors.lastname: Zhang
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Yan-chun
authors.initials: YC
authors.lastname: Ji
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Ping-ping
authors.initials: PP
authors.lastname: Jiang
authors.affiliation: Institute of Genetics, Zhejiang University and Department of Genetics, Zhejiang University School of Medicine, Hangzhou 310058, China.
authors.forename: Min-Xin
authors.initials: MX
authors.lastname: Guan
mesh_terms: D045744:Cell Line, Tumor; D004272:DNA, Mitochondrial / Q000235:genetics*; D018628:Gene Dosage; D006801:Humans; D008214:Lymphocytes / Q000378:metabolism*; D008928:Mitochondria / Q000378:metabolism; D028361:Mitochondrial Diseases / Q000209:etiology / Q000235:genetics*; D009154:Mutation; D010101:Oxygen Consumption
chemical_list: D004272:DNA, Mitochondrial