ACOUSTIC PROCESSES FOR TRANSFECTION AND TRANSDUCTION
Methods for introducing foreign nucleic acids into cells, such as by performing transfection/transduction, using acoustic processes are disclosed herein. The foreign DNA/RNA and the cells are co-located in a multi-dimensional acoustic standing wave, or are co-located by acoustic streaming.
1 . A method for introducing foreign nucleic acids into cells, comprising:
placing the cells and nucleic acids in an acoustophoretic device comprising:
an acoustic chamber in which the cells and the nucleic acids are placed; and
an ultrasonic transducer including a piezoelectric material that can be driven to create an acoustic standing wave in the acoustic chamber; and
driving the ultrasonic transducer to create the multi-dimensional acoustic standing wave;
wherein at least the cells are retained by the acoustic standing wave, the nucleic acids being co-located with the cells to permit introduction of the foreign nucleic acids into the cells.
2 . The method of claim 1 , wherein the nucleic acids are in a viral vector.
3 . The method of claim 1 , further comprising opening pores in cell membranes of the cells prior to co-locating the cells with the nucleic acids.
4 . The method of claim 3 , wherein the pores are opened by electroporation, sonoporation, or by exposure to calcium phosphate.
5 . The method of claim 1 , wherein the acoustophoretic device further comprises a recirculation loop coupled to the acoustic chamber; and one or more of the cells or the nucleic acids are recirculated through the acoustic chamber.
6 . The method of claim 1 , wherein the cells are Chinese hamster ovary (CHO) cells, NS0 hybridoma cells, baby hamster kidney (BHK) cells, human cells, regulatory T-cells, helper T-cells, cytotoxic T-cells, memory T-cells, effector T-cells, gamma delta T-cells, Jurkat T-cells, CAR-T cells, B cells, or NK cells, peripheral blood mononuclear cells (PBMCs), algae, plant cells, or bacteria.
7 . The method of claim 1 , wherein the acoustic standing wave is a multi-dimensional acoustic standing wave, a planar standing wave, or a combination of a multi-dimensional acoustic standing wave and a planar standing wave.
8 . The method of claim 1 , wherein the ultrasonic transducer is driven at a frequency of about 0.5 MHz to about 20 MHz.
9 . The method of claim 1 , wherein the frequency of the acoustic standing wave is varied in a sweep pattern to move the cells relative to the nucleic acids.
10 . A method for causing transduction of cells, comprising:
placing the cells and a viral vector comprising nucleic acids in an acoustophoretic device comprising:
an acoustic chamber in which the cells and the viral vector are placed; and
an ultrasonic transducer including a piezoelectric material that can be driven to create an acoustic standing wave in the acoustic chamber; and
driving the ultrasonic transducer to create the multi-dimensional acoustic standing wave;
wherein the cells and the viral vector are co-located by the acoustic standing wave to permit transduction of the cells.
11 . The method of claim 10 , wherein the cells and the viral vector are suspended in a fluid.
12 . The method of claim 10 , wherein the cells are Chinese hamster ovary (CHO) cells, NS0 hybridoma cells, baby hamster kidney (BHK) cells, human cells, regulatory T-cells, helper T-cells, cytotoxic T-cells, memory T-cells, effector T-cells, gamma delta T-cells, Jurkat T-cells, CAR-T cells, B cells, or NK cells, peripheral blood mononuclear cells (PBMCs), algae, plant cells, or bacteria.
13 . The method of claim 10 , wherein the acoustophoretic device further comprises a recirculation loop coupled to the acoustic chamber; and one or more of the cells or the nucleic acids are recirculated through the acoustic chamber.
14 . The method of claim 10 , wherein the ultrasonic transducer is driven at a frequency of about 0.5 MHz to about 20 MHz.
15 . The method of claim 10 , wherein the frequency of the acoustic standing wave is varied in a sweep pattern to move the cells relative to the viral vector.
16 . A method for causing transfection of cells, comprising:
opening pores in cell membranes of the cells;
placing the cells and nucleic acids in an acoustophoretic device comprising:
an acoustic chamber in which the cells and the nucleic acids are placed; and
an ultrasonic transducer including a piezoelectric material that can be driven to create an acoustic standing wave in the acoustic chamber; and
driving the ultrasonic transducer to create the acoustic standing wave;
wherein the cells and the nucleic acids are co-located by the acoustic standing wave to cause transfection of the cells.
17 . The method of claim 16 , wherein the pores are opened by electroporation, sonoporation, or by exposure to calcium phosphate.
18 . The method of claim 16 , wherein the pores are opened before or after the cells are placed in the acoustophoretic device.
19 . The method of claim 16 , wherein the acoustophoretic device further comprises a recirculation loop coupled to the acoustic chamber; and one or more of the cells or the nucleic acids are recirculated through the acoustic chamber.
20 . The method of claim 16 , wherein the cells are Chinese hamster ovary (CHO) cells, NS0 hybridoma cells, baby hamster kidney (BHK) cells, human cells, regulatory T-cells, helper T-cells, cytotoxic T-cells, memory T-cells, effector T-cells, gamma delta T-cells, Jurkat T-cells, CAR-T cells, B cells, or NK cells, peripheral blood mononuclear cells (PBMCs), algae, plant cells, or bacteria.
21 . The method of claim 16 , wherein the acoustic standing wave is a multi-dimensional acoustic standing wave, a planar standing wave, or a combination of a multi-dimensional acoustic standing wave and a planar standing wave.
22 . The method of claim 16 , wherein the ultrasonic transducer is driven at a frequency of about 0.5 MHz to about 20 MHz.
23 . The method of claim 16 , wherein the frequency of the acoustic standing wave is varied in a sweep pattern to move the cells relative to the nucleic acids.
24 . The cells produced by the method of claim 1 .
25 . The cells produced by the method of claim 10 .
26 . The cells produced by the method of claim 16 .