Cell manipulation technology using nanoneedle arrays

To manipulate a large number of cells simultaneously, we have established a process to fabricate nanoneedle array (NNA) by deep reactive ion etching to silicon wafer. As with the AFM cantilever-type nanoneedles, the insertion damage of NNA is extremely small and the isolated stem cells can be differentiated into cell of interest correctly.

Tens of thousands of nanoneedles with a diameter of 200 nm and a length of over 30 µm are regularly arranged on a 5 mm square silicon chip.

Highly efficient and safe genome editing is achieved by adsorbing Cas9/gRNA complex onto the nanoneedle array and inserting it into cells. Genome editing efficiency of 16% has been achieved.

Antibody-modified nanoneedle arrays are simultaneously inserted into cell arrays in which cell adhesion forces are equally adjusted to about 10 nN, and the cells are pulled up by fishing force and collected on a strongly adherent substrate. We have succeeded in isolating human iPS-derived neural stem cells.