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The cell membrane is a selectively permeable barrier between a cell and its environment, regulating passage of material into and out of the cell.

Membrane transport is fundamental to the intrinsic functioning of the cell with several natural mechanisms (e.g., passive diffusion, active and co-transport, and endocytosis/exocytosis) permitting cellular uptake and secretion of small and large molecules.

In practice, this unique capability is negated by the low throughput of the method.

Field-mediated membrane poration has supplanted chemical methods in many delivery applications, particularly those involving nongene target molecules and primary cells.

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This is not true of larger macromolecules, which require alternative means to enter the cell interior.

Ideal delivery systems also protect materials from cytoplasmic degradation, convey materials to a target location, and facilitate action on that target.

Acoustic Shear Poration (ASP) allows passive cytoplasmic delivery of small to large nongene macromolecules into established and primary cells at greater than 75% efficiency.

This facile parallelization and scale up are crucial to therapeutic applications and cell-based biomanufacturing, where sample sizes can exceed billions of cells.

Delivery of macromolecules such as nucleic acids to primary cells ex vivo is a critical component of many new cell-based therapies such as adoptive T-cell immunotherapy.

Addition of an electrophoretic action enables active transport of target DNA molecules to substantially augment transfection efficiency of passive mechanoporation/diffusive delivery without affecting viability.

This two-stage poration/insertion method preserves the compelling flexibility of shear-based delivery, yet substantially enhances capabilities for active transport and transfection of plasmid DNA.

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Converging nozzle-like channels were used to achieve DNA transfection via focused acoustic pressure driven cell mechanoporation. have demonstrated the insensitivity of these methods to cell type and target molecule, providing additional evidence to support their potential as a universal route to in vitro and ex vivo delivery.

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