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. Author manuscript; available in PMC: 2015 Aug 1.
Published in final edited form as: Biochim Biophys Acta. 2013 Sep 23;1842(8):1219–1231. doi: 10.1016/j.bbadis.2013.09.010

Figure 2. The cybrid technique.

Figure 2

An expandable cell line is treated with enough ethidium bromide to interfere with mtDNA replication, but not enough to prevent nuclear DNA replication. This depletes the cell line's endogenous mtDNA (shown by the absence of the red mtDNA circle within the mitochondrion in the top row), removes its mtDNA-encoded subunits (shown by the absence of the red rectangles within the mitochondrion in the top row), and generates a respiration-incompetent cell called a ρ0 cell. Incubating ρ0 cells with platelets in the presence of polyethylene glycol (PEG) allows platelet and ρ0 cell cytosols to mix. Platelet mitochondria contain mtDNA (shown as a green circle within the mitochondrion in the bottom row), which populates the ρ0 cell mitochondria, generates mtDNA-encoded respiratory chain subunits (shown as green rectangles within the mitochondrion in the bottom row), and restores respiratory competence. The new cybrid cell now contains mtDNA from the platelet donor, mtDNA-encoded respiratory chain subunits that match those of the platelet donor, its original nuclear DNA, and its original nuclear-encoded respiratory chain subunits.