Abstract: The present disclosure provides a microfluidic device for buffer exchange of suspended particles comprising a channel having a first inlet for a first fluid containing suspended particles and a second inlet for a second fluid. The device includes an angled ridge positioned within the channel at 5 degrees or less relative to a channel wall parallel to flow direction, extending partially across channel width and configured to deflect particles from the first fluid into the second fluid while allowing fluids to pass through a ridge gap. A gutter region adjacent to the angled ridge distal end allows particles to pass through. An outlet collects particles transferred into the second fluid.

Abstract: The present disclosure provides methods and systems for cell processing, including delivery of imaging agents into cells. The methods and systems may comprise the use of a microfluidic device. The microfluidic device may comprise a channel comprising a compressive element. The compressive element may be configured to reduce a volume of the cell and facilitate the formation of one or more transient pores in a cell membrane of the cell. The one or more pores may permit one or more imaging agents to enter the cell. Also provided are modified cells produced using the disclosed methods and systems and methods of imaging the modified cells in a subject.

Abstract: Described herein are high-throughput, high-volume mechanoporation systems and methods for intracellular payload delivery, enabling scalable and cost-effective cell therapy manufacturing. The apparatus includes a pre-processing media reservoir with a media cavity and a processing stack comprising two end gaskets and a processing unit. The processing unit features inlet and outlet slots, processing channels fluidically connecting them, and ridges forming gaps within the channels. The inlet slots align with gasket slots in the end gasket, allowing media to flow toward the ridges. When pressure is applied to the media reservoir, the media is forced through the gaps in the processing channels, inducing mechanoporation of cells. This system automates and streamlines cell engineering, improving throughput, consistency, and scalability while reducing costs, aligning with the needs of advanced cell therapy production.

Abstract: Described herein are methods and systems for mechanoporation-based high-throughput payload delivery into biological cells. For example, one system can process at least 1 billion cells per minute or at least 25 billion cells per minute, which is substantially greater than conventional methods. A cell processing apparatus comprises a processing assembly formed by stacking multiple processing components. Each processing component comprises channels, which may be used for filtration, mechanoporation, and/or separation of cells in the cell media. This functionality depends on the configuration of each channel. For example, each channel comprises one or more ridges such that each ridge forms a processing gap with an adjacent one of the processing components. The ridges may extend to the side walls or form a bypass gap with the wall. The processing gaps can be specially configured to compress cells as the cells pass through these gaps thereby initiating the mechanoporation process.

Abstract: Described herein are methods and systems for cell processing or, more specifically, for introducing various payloads into cells. These methods and systems use a mechanoporation approach in which cells are rapidly compressed and then released to relax while absorbing the payload. More specifically, these methods and systems enable high-throughput mechanoporation with various clogging mitigation features. A cell processing apparatus comprises a shell with an inner shell cylindrical surface, a core with an outer core cylindrical surface, and ridges, supported on and protruding away from one of the inner shell cylindrical surface and the outer core cylindrical surface. The core is disposed inside the shell. The outer core cylindrical surface is concentric with the inner shell cylindrical surface. Each of the ridges forms a ridge gap with the other one of the inner shell cylindrical surface and the outer core cylindrical surface.

Abstract: Microfluidic devices for cell sorting or cell fractionation are disclosed. A microfluidic device can comprise one or more inlets, a first wall and a second wall, and two or more outlets. The first and second walls can be substantially planar to each other and the first wall having can have a plurality of ridges protruding from the first wall and defining a compression gap between the ridge and a surface of the second wall. The microfluidic device can also be a cell sorting device for sorting a plurality of cells based on one or more biophysical cellular properties including size, elasticity, viscosity, and/or viscoelasticity wherein the cells are subjected to one or more compressions due to the compression gap. Also disclosed are methods for cell sorting based on a variety of biophysical cellular properties.

Abstract: Described herein are methods and systems for mechanoporation-based high-throughput payload delivery into biological cells. For example, one system can process at least 1 billion cells per minute or at least 25 billion cells per minute, which is substantially greater than conventional methods. A cell processing apparatus comprises a processing assembly formed by stacking multiple processing components. Each processing component comprises channels, which may be used for filtration, mechanoporation, and/or separation of cells in the cell media. This functionality depends on the configuration of each channel. For example, each channel comprises one or more ridges such that each ridge forms a processing gap with an adjacent one of the processing components. The ridges may extend to the side walls or form a bypass gap with the wall. The processing gaps can be specially configured to compress cells as the cells pass through these gaps thereby initiating the mechanoporation process.

Abstract: Described herein are methods and systems for mechanoporation-based high-throughput payload delivery into biological cells. For example, one system can process at least 1 billion cells per minute or at least 25 billion cells per minute, which is substantially greater than conventional methods. A cell processing apparatus comprises a processing assembly formed by stacking multiple processing components. Each processing component comprises channels, which may be used for filtration, mechanoporation, and/or separation of cells in the cell media. This functionality depends on the configuration of each channel. For example, each channel comprises one or more ridges such that each ridge forms a processing gap with an adjacent one of the processing components. The ridges may extend to the side walls or form a bypass gap with the wall. The processing gaps can be specially configured to compress cells as the cells pass through these gaps thereby initiating the mechanoporation process.

Abstract: Described herein are methods and systems for mechanoporation-based high-throughput payload delivery into biological cells. For example, one system can process at least 1 billion cells per minute or at least 25 billion cells per minute, which is substantially greater than conventional methods. A cell processing apparatus comprises a processing assembly formed by stacking multiple processing components. Each processing component comprises channels, which may be used for filtration, mechanoporation, and/or separation of cells in the cell media. This functionality depends on the configuration of each channel. For example, each channel comprises one or more ridges such that each ridge forms a processing gap with an adjacent one of the processing components. The ridges may extend to the side walls or form a bypass gap with the wall. The processing gaps can be specially configured to compress cells as the cells pass through these gaps thereby initiating the mechanoporation process.

Abstract: A microchannel for processing cells by compression of the cells including an inlet, ridges and an outlet. Each ridge including a compressive surface and a cell adhesion entity. The outlet configured to remove at least one of a first portion of the cells and a second portion of the cells from the microchannel. Each ridge oriented at an angle of from 25 degrees to 70 degrees relative to a center axis of the microchannel. The cell adhesion entity configured such that the first portion of the cells has a first adhesion property relative to the cell adhesion entity to follow a first trajectory through the microchannel. The cell adhesion entity further configured such that the second portion of the cells has a second adhesion property relative to the cell adhesion entity to follow a second trajectory through the microchannel. The first trajectory is different from the second trajectory.

Abstract: The present disclosure provides methods and systems for cell processing, including delivery of imaging agents into cells. The methods and systems may comprise the use of a microfluidic device. The microfluidic device may comprise a channel comprising a compressive element. The compressive element may be configured to reduce a volume of the cell and facilitate the formation of one or more transient pores in a cell membrane of the cell. The one or more pores may permit one or more imaging agents to enter the cell. Also provided are modified cells produced using the disclosed methods and systems and methods of imaging the modified cells in a subject.

Abstract: A microchannel for processing cells by compression of the cells including an inlet, ridges and an outlet. Each ridge including a compressive surface and a cell adhesion entity. The outlet configured to remove at least one of a first portion of the cells and a second portion of the cells from the microchannel. Each ridge oriented at an angle of from 25 degrees to 70 degrees relative to a center axis of the microchannel. The cell adhesion entity configured such that the first portion of the cells has a first adhesion property relative to the cell adhesion entity to follow a first trajectory through the microchannel. The cell adhesion entity further configured such that the second portion of the cells has a second adhesion property relative to the cell adhesion entity to follow a second trajectory through the microchannel. The first trajectory is different from the second trajectory.

Abstract: The present disclosure provides methods and systems for cell processing, including delivery of substances into cells. The methods and systems may comprise the use of a microfluidic device. The microfluidic device may comprise a channel comprising a compressive element. The compressive element may be configured to reduce a volume of the cell and facilitate the formation of one or more transient pores in a cell membrane of the cell. The one or more pores may permit one or more substances such as therapeutic or gene-editing reagents to enter the cell. Also provided are modified cells produced using the disclosed methods and systems.

Abstract: Described herein are methods and systems for cell processing or, more specifically, for introducing various payloads into cells. These methods and systems use a mechanoporation approach in which cells are rapidly compressed and then released to relax while absorbing the payload. More specifically, these methods and systems enable high-throughput mechanoporation with various clogging mitigation features. A cell processing apparatus comprises a shell with an inner shell cylindrical surface, a core with an outer core cylindrical surface, and ridges, supported on and protruding away from one of the inner shell cylindrical surface and the outer core cylindrical surface. The core is disposed inside the shell. The outer core cylindrical surface is concentric with the inner shell cylindrical surface. Each of the ridges forms a ridge gap with the other one of the inner shell cylindrical surface and the outer core cylindrical surface.

Abstract: Embodiments of the present disclosure can include a method comprising: providing a plurality of cells to a microchannel, the microchannel coated in at least one cell adhesion entity and comprising a compressive surface and a first outlet, the compressive surface defining a compression gap, flowing the plurality of cells through the microchannel, wherein the flowing comprises: compressing the plurality of cells underneath the compressive surface; and exposing the plurality of cells to the at least one cell adhesion entity, wherein the exposing causes a first portion of the cells having a first adhesion property to temporarily bind to the cell adhesion entity; and collecting the first portion of cells at the first outlet; wherein the compression gap has a height of from 75% to 95% an average diameter of the plurality of cells.

Abstract: Embodiments of the present disclosure can include a method for convective intracellular delivery including providing cells and molecules to a microchannel having compressive surfaces, wherein the compressive surfaces define compression gaps having a height of from 20 and 80% of the average cell diameter, and a plurality of relaxation spaces disposed between the compressive surfaces, flowing the cell medium through the microchannel, wherein as the cell medium flows through the microchannel, the plurality of cells undergo a convective intracellular delivery process comprising: compressing the plurality of cells, wherein the compressing causes the plurality of cells to undergo a loss in intracellular volume Vloss, and passing the plurality of cells to a first relaxation space, wherein the plurality of cells undergo a gain in volume Vgain and absorb a portion of the plurality of molecules.

Abstract: The present disclosure provides systems and methods for intracellular delivery. The systems and methods may comprise the use of a cell processing apparatus which may comprise a plurality of compression elements such as ridges. The intracellular delivery may be caused by rapid compression of cells, which may result in a reduction of a cell volume. The compression may occur while the cells pass through gaps formed by at least a subset of the ridges. The cell processing apparatus may further comprise one or more recovery spaces which are positioned between adjacent ridges. The cells may recover at least a portion of the reduced cell volume by absorbing media and/or reagents surrounding the cells while flowing through the recovery spaces. The ridges may also divert less compressible cells into a diversion channel, thereby sorting the cells based on various cell properties and/or preventing clogging within the apparatus.

Abstract: Embodiments of the present disclosure can include a method for convective intracellular delivery including providing cells and molecules to a microchannel having compressive surfaces, wherein the compressive surfaces define compression gaps having a height of from 20 and 80% of the average cell diameter; and a plurality of relaxation spaces disposed between the compressive surfaces; flowing the cell medium through the microchannel, wherein as the cell medium flows through the microchannel, the plurality of cells undergo a convective intracellular delivery process comprising: compressing the plurality of cells, wherein the compressing causes the plurality of cells to undergo a loss in intracellular volume (Vloss); and passing the plurality of cells to a first relaxation space, wherein the plurality of cells undergo a gain in volume (Vgain) and absorb a portion of the plurality of molecules.

Abstract: The present disclosure provides systems and methods for intracellular delivery. The systems and methods may comprise the use of a cell processing apparatus which may comprise a plurality of compression elements such as ridges. The intracellular delivery may be caused by rapid compression of cells, which may result in a reduction of a cell volume. The compression may occur while the cells pass through gaps formed by at least a subset of the ridges. The cell processing apparatus may further comprise one or more recovery spaces which are positioned between adjacent ridges. The cells may recover at least a portion of the reduced cell volume by absorbing media and/or reagents surrounding the cells while flowing through the recovery spaces. The ridges may also divert less compressible cells into a diversion channel, thereby sorting the cells based on various cell properties and/or preventing clogging within the apparatus.

Abstract: Microfluidic devices for cell sorting or cell fractionation are disclosed. A microfluidic device can comprise one or more inlets, a first wall and a second wall, and two or more outlets. The first and second walls can be substantially planar to each other and the first wall having can have a plurality of ridges protruding from the first wall and defining a compression gap between the ridge and a surface of the second wall. The microfluidic device can also be a cell sorting device for sorting a plurality of cells based on one or more biophysical cellular properties including size, elasticity, viscosity, and/or viscoelasticity wherein the cells are subjected to one or more compressions due to the compression gap. Also disclosed are methods for cell sorting based on a variety of biophysical cellular properties.