US 12,030,052 B2
Microfluidic delivery method utilizing an electric field
Ryan Pawell, San Francisco, CA (US); Amy Twite, Berkeley, CA (US); Geoff Facer, Redwood City, CA (US); Katherine Lau, Alameda, CA (US); Adrian Lievano, San Francisco, CA (US); and Julyana Acevedo, Berkeley, CA (US)
Filed by Indee. Pty. Ltd., Sydney (AU)
Filed on Jul. 5, 2021, as Appl. No. 17/367,619.
Application 17/367,619 is a division of application No. 16/474,996, granted, now 11,052,394, previously published as PCT/AU2018/051190, filed on Nov. 2, 2018.
Claims priority of provisional application 62/580,922, filed on Nov. 2, 2017.
Prior Publication US 2021/0339252 A1, Nov. 4, 2021
Int. Cl. B01L 3/00 (2006.01); A61K 35/15 (2015.01); A61K 35/17 (2015.01); C12N 13/00 (2006.01); C12N 15/64 (2006.01); C12N 15/87 (2006.01); C12N 15/90 (2006.01)
CPC B01L 3/502761 (2013.01) [A61K 35/15 (2013.01); A61K 35/17 (2013.01); B01L 3/502715 (2013.01); B01L 3/502746 (2013.01); C12N 13/00 (2013.01); C12N 15/64 (2013.01); C12N 15/87 (2013.01); C12N 15/90 (2013.01); B01L 2200/0647 (2013.01); B01L 2400/0415 (2013.01); B01L 2400/082 (2013.01)] 11 Claims
 
1. A method for introducing exogenous material into a cell, comprising:
exposing the cell in suspension with the exogenous material to a pressure change under unsteady flow conditions to temporarily permeabilise a cell membrane without the cell becoming lysed; and
exposing the cell to an electrical field to introduce the exogenous material into the cell while the membrane is permeabilised, the electric field being generated at an electric field strength insufficient to adversely perturb the cell state.
 
10. A method for introducing exogenous material into a cell, comprising:
exposing the cell in suspension with the exogenous material to a pressure change under unsteady flow conditions to temporarily permeabilise a cell membrane without the cell becoming lysed; and
exposing the cell to an electrical field to introduce the exogenous material into the cell while the membrane is permeabilized, the electric field including an oscillating component and an offset insufficient to adversely perturb the cell state.
 
11. A method for introducing exogenous material into a cell, comprising:
exposing the cell in suspension with the exogenous material to a pressure change under unsteady flow conditions to temporarily permeabilise a cell membrane without the cell becoming lysed; and
exposing the cell to an electrical field to introduce the exogenous material into the cell while the membrane is permeabilized, the electric field being generated at a frequency insufficient to adversely perturb the cell state.