Transmitochondrial technology in animal cells

Abstract

Studies of vertebrate mitochondrial DNA (mtDNA) maintenance and function have relied heavily on somatic cell experimentation in culture as it is difficult to manipulate mtDNA sequences in animal cells. Initial patterns of mitochondrial segregation and species-specific compatibility have been performed using somatic hybrid and cybrid cells. These studies became easier to interpret with the development of cell lines devoid of mtDNA. This technology has also been used to study evolutionary interactions between nuclear and mitochondrial genomes by closely related species. Different tissue types and techniques have been utilized for the generation of transmitochondrial cell lines in animals. Basically, a cell line devoid of mtDNA or with poisoned mitochondria function as the nuclear donor, whereas enucleated or fragmented cells function as the mtDNA donor. Upon cell membrane fusion and appropriate selection, transmitochondrial cells are generated. This chapter describes current protocols used to generate transmitochondrial animal cell lines in culture.