A Novel Cell Manipulation Device for Cytoplasmic Transplantation

We present novel cell manipulation devices for cytoplasmic transplantation without mixing of genomes. The device consists of two channels separated by an insulating wall on which an orifice is fabricated. The orifice was utilized for a high-yield electrofusion between donor and recipient cells, and physical removal of the nucleus belonging to the donor. A donor and a recipient cell for the transplantation were first brought into contact through an orifice by dielectrophoresis (DEP). Then, voltage pulse was applied to induce cell membrane breakdown. The electric field lines are constricted at an orifice so that the membrane breakdown occurs exclusively at an orifice, inducing cell fusion at an orifice. After the cell fusion, the fusant was taken-out toward the recipient side by applying a pressure difference. Because the orifice was small enough to prevent the donor nucleus from being taken-out together, only the cytoplasm was transplanted. We also demonstrate feasibility of parallel processing of the transplantations using a device which has an array of orifice on an insulating sheet. A large number of cells were successfully fusioned in parallel by using the orifice-sheet. The parallel cytoplasmic transplantation would be realized by making the size of orifices smaller.