FIG. 3.

The cybrid technique. mtDNA-depleted (ρ0) cells line and non-nucleated cytoplasts or platelets are mixed in the presence of a plasma membrane-disrupting detergent. After the detergent is diluted, membrane reconstitution generates some cells that contain a nucleus from a ρ0 cell, ρ0 cytosolic contents, and cytoplast cytosolic contents. Only cybrid cells that contain cytoplast mtDNA can go on to produce functional ETCs and survive a subsequent selection process. After the selection process is complete, the resulting cybrid cell line is used for biochemical and molecular analyses. Because cybrid cell lines created from a common ρ0 cell nuclear background have equivalent nuclear genes, and the cell expansion environment is equivalent between cell lines, biochemical or molecular differences between lines are expected to reflect differences between their mtDNA content.