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US 12,628,820 B2Granted
Systems and methods for preservation of cells
AssigneeUniversity of Louisville Research Foundation, Inc.
InventorsMichael Menze, Jonathan Kopechek, Brett Janis
Life of this patent
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Abstract
Provided herein are systems and methods for molecule delivery into cells. The method may include steps of forming a solution comprising an amount of microbubbles, an amount of soluble molecules, and one or more cells, flowing the solution through a microfluidic device, and sonicating the solution for a period of time sufficient to form pores in the one or more cells. In particular, such methods may be used for preservation of cells through delivery of a preservation agent.
Claims15
A method of using sonication to deliver soluble molecules into a plurality of untreated cells to produce treated cells, the method comprising:
- forming a solution comprising an amount of naked positively charged lipid coated microbubbles, an amount of soluble molecules, and a plurality of untreated cells;
- providing a microfluidic device comprising at least a first channel;
- providing an ultrasound device capable of emitting ultrasound waves;
- positioning at least a portion of the first channel in proximity to the ultrasound device;
- passing the solution through the first channel at a sustained predetermined flow rate;
- as the solution passes through the portion of the first channel positioned in proximity to the ultrasound device at the predetermined flow rate, sonoporating a membrane of at least one of the plurality of untreated cells by emitting ultrasound waves from the ultrasound device for a period of time sufficient for the naked positively charged lipid coated microbubbles in the solution to cause a microjetting effect within the fluid surrounding the plurality of untreated cells, forming transient pores in the membrane of at least one of the plurality of untreated cells;
- and
- wherein the microjetting effect within the solution is sufficient to cause at least a portion of the amount of soluble molecules in the solution to be delivered through the porous membrane of the at least one untreated cell to form at least one treated cell, and wherein the plurality of untreated cells are red blood cells.
5 more elements
A method of using sonication to preserve a plurality of red blood cells, the method comprising:
- forming a solution comprising an amount of naked positively charged lipid coated microbubbles, an amount of preservation agent, and a plurality of unpreserved red blood cells;
- providing a microfluidic device comprising at least a first channel;
- providing an ultrasound device capable of emitting ultrasound waves;
- positioning at least a portion of the first channel in proximity to the ultrasound device;
- passing the solution through the first channel at a sustained predetermined flow rate;
- as the solution passes through the portion of the first channel positioned in proximity to the ultrasound device at the predetermined flow rate, sonoporating a membrane of at least one of the plurality of unpreserved red blood cells by emitting ultrasound waves from the ultrasound device for a period of time sufficient for the naked positively charged lipid coated microbubbles in the solution to cause a microjetting effect within the fluid surrounding the plurality of unpreserved red blood cells, forming transient pores in the membrane of at least one of the plurality of unpreserved red blood cells;
- and
- wherein the microjetting effect within the solution is sufficient to cause at least a portion of the amount of preservation agent in the solution to be delivered through the porous membrane of the at least one unpreserved red blood cell to form at least one preserved red blood cell.
5 more elements
A method of using sonoporation to deliver soluble molecules into untreated cells, the method comprising:
- passing a solution comprising an amount of naked positively charged lipid coated microbubbles, an amount of soluble molecules, and one or more untreated cells through a microchannel having a shape that causes centrifugal force to be applied to the solution;
- providing an ultrasound device capable of emitting ultrasound waves;
- positioning at least a portion of the microchannel in proximity to the ultrasound device;
- passing the solution through the microchannel at a sustained predetermined flow rate;
- as the solution passes through the portion of the microchannel positioned in proximity to the ultrasound device at the predetermined flow rate, sonoporating a membrane of at least one of the untreated cells by emitting ultrasound waves from the ultrasound device for a period of time sufficient for the naked positively charged lipid coated microbubbles in the solution to cause a microjetting effect within the fluid surrounding the one or more untreated cells, forming transient pores in the membrane of at least one of the untreated cells;
- and
- wherein the microjetting effect within the solution is sufficient to cause at least a portion of the amount of soluble molecules in the solution to be delivered through the porous membrane of the at least one untreated cell to form at least one treated cell, and wherein the one or more untreated cells are red blood cells.
4 more elements
Dependencies are inferred from the claim text.