[27] Isolation of human cell lines lacking mitochondrial DNA

This chapter discusses isolation of human cell lines lacking mitochondrial DNA. An understanding of the molecular genetic mechanisms by which these mutations act can provide insights into the etiology, pathogenesis, and ultimately the treatment of these diseases, and may enhance the knowledge of mitochondrial biogenesis. For investigating nucleomitochondrial interactions and the molecular pathogenetic mechanisms of mtDNA mutations, it is useful to manipulate the mtDNA complement of a cell, move mitochondria from one cellular environment to another, or introduce new genes into mitochondria. As an initial step towards these goals, there is an isolation human cell lines that completely lack mtDNA (ρ0 cell lines). The chapter describes the theory and the methods utilized to isolate such cell lines.