Cartridge and device for use in a system for intracellular payload delivery - Patents.com

Abstract

A device for facilitating delivery of a payload to cells in a cell suspension, the device comprising a container and a cartridge, the cartridge configured to couple to an opening of the container when in an assembled position relative to the container, the cartridge including an input reservoir configured to receive the cell suspension, the cartridge including a housing configured to house a constriction-containing element, the constriction-containing element including a constriction configured to perturb a membrane of cells in the cell suspension to facilitate delivery of the payload to the cells, the housing portion fluidly coupled to the input reservoir, thereby allowing the cell suspension to flow from the input reservoir to the housing portion, and the housing portion fluidly coupled to a cartridge outlet, thereby allowing the cell suspension to flow from the housing portion into an interior cavity of the container.

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to and the benefit of U.S. Provisional Patent Application No. 63 / 346,634, filed May 27, 2022, the entire contents of which are hereby incorporated by reference for all purposes.

[0002] The present disclosure relates to systems for delivering a payload into a cell, and more particularly to cartridges and devices that hold a constriction-containing element having a constriction channel or pore, the use of which in the system causes a perturbation of the cell membrane, allowing the payload to pass through the cell membrane. [Background technology]

[0003] Controlling the delivery of various materials into cells is important in the evolving medical field of cell therapy. For example, various research and therapeutic applications may involve delivering peptides, nucleic acids, proteins, small molecules, and nanomaterials into cells through cell membranes. As discussed in WO2013059343, WO2015023982, PCT / US2015 / 058489, PCT / US2015 / 060689, and PCT / US2016 / 13113, constricted microfluidic channels may be used to deliver compounds and other payloads into cells. As disclosed in PCT / US18 / 66295, benchtop laboratory and / or clinical systems may be configured to force a cell suspension through a cartridge that contains one or more constriction-containing elements (e.g., parts, parts, devices, or components such as microfluidic chips or filters) having constricted channels or constricted holes to cause perturbations in the membranes of cells in the cell suspension. Summary of the Invention

[0004] As explained above, some systems for intracellular payload delivery include a cartridge configured to house one or more constriction-containing elements (e.g., microfluidic chips or filters) with constriction channels or constriction holes to induce perturbations in the membranes of cells in the cell suspension as the cell suspension flows through the cartridge. However, the cartridges of these systems may leave room for improvement. For example, such systems do not allow for a spatially efficient and concise integration of the cartridge with an input reservoir (e.g., a reservoir for holding fluid before the fluid passes through the cartridge) and an output reservoir (e.g., a reservoir for holding fluid after the fluid passes through the cartridge). Such systems rely, for example, on the use of flexible tubing to connect the cartridge to the input reservoir and / or the output reservoir, which may be provided, for example, in the form of a flexible bag. These systems are not configured, for example, with respect to space efficiency, durability, simplicity, compatibility with existing laboratory equipment, and / or compatibility with robotic control systems.

[0005] Thus, there is a need for a cartridge that has improved geometry, improved space efficiency, improved durability, improved simplicity, improved compatibility with existing laboratory equipment, and / or improved compatibility with robotic control systems. The systems, methods, and techniques disclosed herein may address one or more of the needs identified above.

[0006] Disclosed herein are systems and devices for use in delivering payloads into cells. The devices include a cartridge configured to receive and hold a constriction-containing element, such as a microfluidic chip or a microfluidic filter. Unlike cartridge systems that may be configured to be attached to an input reservoir by a flexible tube or other fluidics component, the cartridges disclosed herein may include an input reservoir that is physically integrated with the cartridge itself. The input reservoir may be a reservoir configured to contain a cell suspension before the cell suspension flows through the constriction-containing element. The cartridges disclosed herein may be provided in the form of a cap device that is coupled to and configured to cover the opening of a vessel, such as a test tube, culture tube, sample tube, or centrifuge tube. The use of an integrated input reservoir facilitates integration with existing laboratory devices, such as readily available pressure sources. Additionally, the timing and volume reduction potential of using the embodiments described herein may be particularly useful, for example, in research use cases where volumes are small and consumables can be used quickly with multiple small volume experiments. A vessel (e.g., a tube) to which the cartridge may be attached as a cap may function as an output reservoir to contain the cell suspension after processing through the constriction-containing element. In some embodiments, when the cartridge is in an attached position with the vessel (e.g., a tube), a portion of the cartridge may be disposed inside the vessel's internal cavity, for example by projecting downwardly into the interior of a test tube. This configuration may facilitate retrieval of the tubular vessel, further facilitating its use in a research setting.

[0007] Thus, when the cartridge is attached to the open end of a vessel, such as a laboratory tube, the device formed by the combination of the cartridge and vessel may include an input reservoir, a housing portion configured to receive and hold a constriction-containing element, and an output reservoir provided in the form of a vessel, such as a laboratory tube. All three components may be in fluid communication with each other, allowing the cell suspension to flow from the input reservoir to the housing portion (and into and out of the constriction-containing element contained in the housing portion) and then from the housing portion to the vessel (e.g., tube). After the cell suspension has flowed into the vessel (e.g., tube), the cartridge may be removed from the opening of the vessel and the vessel (containing the processed cell suspension) may be used for further processing of the cell suspension, may be capped, and / or may be stored in any suitable manner. If the vessel is provided in the form of a standard laboratory tube, the tube may be easily used for downstream processing with one or more standard-compatible additional devices, caps, and / or storage devices.

[0008] Some embodiments are directed to a device for facilitating delivery of a payload to cells in a cell suspension, the device comprising a container including an internal cavity and an opening, and a cartridge, the cartridge configured to couple to the opening of the container when in an assembled position relative to the container. The cartridge includes an input reservoir configured to receive the cell suspension and a storage portion configured to house a constriction-containing element. The constriction-containing element includes a constriction configured to perturb a membrane of cells in the cell suspension to facilitate delivery of the payload to the cells. The storage portion is fluidly coupled to the input reservoir, thereby allowing the cell suspension to flow from the input reservoir to the storage portion, and the storage portion is fluidly coupled directly to a cartridge outlet, thereby allowing the cell suspension to flow from the storage portion into the internal cavity of the container.

[0009] In some embodiments, a portion of the storage portion of the cartridge is disposed inside the internal cavity of the container when the cartridge is in the assembled position relative to the container. In some embodiments, a portion of the input reservoir of the cartridge is disposed inside the internal cavity of the container when the cartridge is in the assembled position relative to the container.

[0010] In some embodiments, the device comprises a cover configured to releasably attach to the storage portion of the cartridge and to hold the constriction-containing element in place when the constriction-containing element is stored in the storage portion. In some embodiments, the cover is configured to be slidably attached and removed from the storage portion of the cartridge. In some embodiments, the cover is configured to slide in a direction perpendicular to the direction in which the layers of constriction-containing elements are stacked on top of each other. In some embodiments, when the cartridge is in an assembled position relative to the container, the cover is configured to be prevented from removal from the storage portion by an inner wall of the container.

[0011] In some embodiments, the cartridge includes a loading reservoir cover that is movable between an open position and a closed position to selectively expose and enclose the interior of the loading reservoir.

[0012] In some embodiments, the cartridge includes a first O-ring disposed against a surface of the input reservoir cover opposite the interior of the input reservoir when the input reservoir cover is in a closed position, hi some embodiments, the cartridge includes a second O-ring disposed between the input reservoir cover and a body portion of the cartridge.

[0013] In some embodiments, the input reservoir cover includes a filter configured to prevent backflow of the cell suspension. In some embodiments, the cartridge has a total fluid throughput of greater than 0.05 L / min. In some embodiments, the cartridge is configured to allow the cell suspension to flow through the cartridge at pressures greater than 100 PSI.

[0014] Some embodiments are directed to a device for facilitating delivery of a payload to cells of a cell suspension. The device comprises a container including an internal cavity and an opening, and a cartridge, the cartridge configured to couple to the opening of the container when in an assembled position relative to the container, the cartridge including an input reservoir configured to receive the cell suspension, and a storage portion. The device comprises a constriction-containing element. The constriction-containing element includes a constriction configured to perturb a membrane of cells of the cell suspension to facilitate delivery of the payload to the cells. The storage portion of the cartridge is fluidly coupled to the input reservoir and the constriction-containing element, which allows the cell suspension to flow from the input reservoir through the storage portion into the constriction-containing element, and allows the cell suspension to flow from the constriction-containing element through the storage portion and into the internal cavity of the container.

[0015] In some embodiments, the device includes a gasket disposed between the containment portion and the constriction-containing element. In some embodiments, the constriction-containing element includes a microfluidic chip that includes a plurality of constrictions that perturb the membrane of the cell. In some embodiments, the constriction-containing element has a footprint of less than 5 mm by 5 mm.

[0016] In some embodiments, a portion of the storage portion of the cartridge is disposed inside the internal cavity of the container when the cartridge is in the assembled position relative to the container. In some embodiments, a portion of the input reservoir of the cartridge is disposed inside the internal cavity of the container when the cartridge is in the assembled position relative to the container.

[0017] In some embodiments, the cartridge further comprises a cover, the cover configured to be releasably attached to the storage portion of the cartridge and to hold the constriction-containing element in place when the constriction-containing element is stored in the storage portion. In some embodiments, the cover is configured to be slidably attached and removed from the storage portion of the cartridge. In some embodiments, the cover is configured to slide in a direction perpendicular to the direction in which the layers of constriction-containing elements are stacked on top of each other. In some embodiments, when the cartridge is in an assembled position relative to the container, the cover is configured to be prevented from being removed from the storage portion by an inner wall of the container.

[0018] In some embodiments, the cartridge includes a loading reservoir cover that is movable between an open position and a closed position to selectively expose and enclose the interior of the loading reservoir.

[0019] In some embodiments, the cartridge includes a first O-ring disposed against a surface of the input reservoir cover opposite the interior of the input reservoir when the input reservoir cover is in a closed position, hi some embodiments, the cartridge includes a second O-ring disposed between the input reservoir cover and a body portion of the cartridge.

[0020] In some embodiments, the input reservoir cover includes a filter configured to prevent backflow of the cell suspension. In some embodiments, the cartridge has a total fluid throughput of greater than 0.05 L / min. In some embodiments, the cartridge is configured to allow the cell suspension to flow through the cartridge at pressures greater than 100 PSI.

[0021] Some embodiments are directed to a cartridge for facilitating delivery of a payload to cells in a cell suspension, the cartridge comprising an input reservoir configured to receive a cell suspension, a coupling portion configured to couple to an opening to an interior cavity of a container, and a storage portion configured to house a constriction-containing element. The constriction-containing element includes a constriction configured to perturb a membrane of cells in the cell suspension to facilitate delivery of the payload to the cells. The storage portion is fluidly coupled to the input reservoir, thereby allowing the cell suspension to flow from the input reservoir to the storage portion, and the storage portion is fluidly coupled to a cartridge outlet, thereby allowing the cell suspension to flow out of the cartridge.

[0022] In some embodiments, a portion of the storage portion of the cartridge is disposed inside the internal cavity of the container when the cartridge is coupled to the container in the assembled position. In some embodiments, a portion of the input reservoir of the cartridge is disposed inside the internal cavity of the container when the cartridge is coupled to the container in the assembled position.

[0023] In some embodiments, the cartridge further comprises a cover, the cover configured to be releasably attached to the storage portion of the cartridge and to hold the constriction-containing element in place when the constriction-containing element is stored in the storage portion. In some embodiments, the cover is configured to be slidably attached and removed from the storage portion of the cartridge. In some embodiments, the cover is configured to slide in a direction perpendicular to the direction in which the layers of constriction-containing elements are stacked on top of each other. In some embodiments, when the cartridge is coupled to the container in an assembled position, the cover is configured to be prevented from being removed from the storage portion by an inner wall of the container.

[0024] In some embodiments, the cartridge includes an input reservoir cover that is movable between an open position and a closed position to selectively expose and enclose the interior of the input reservoir. In some embodiments, the cartridge includes a first O-ring disposed against a surface of the input reservoir cover opposite the interior of the input reservoir when the input reservoir cover is in the closed position. In some embodiments, the cartridge includes a second O-ring disposed between the input reservoir cover and a body portion of the cartridge.

[0025] In some embodiments, the input reservoir cover includes a filter configured to prevent backflow of the cell suspension. In some embodiments, the cartridge has a total fluid throughput of greater than 0.05 L / min. In some embodiments, the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure greater than 100 PSI. In some embodiments, the cartridge has a total fluid throughput of less than 0.5 L / min. In some embodiments, the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure less than 200 PSI.

[0026] In some embodiments, any one or more of the features, characteristics, or elements discussed above with respect to any of the embodiments may be incorporated into any of the other embodiments mentioned above or described elsewhere in this specification. In some embodiments, any one or more of the features, characteristics, or elements discussed elsewhere in this disclosure may be incorporated into any one or more of the embodiments mentioned above. [Brief description of the drawings]

[0027] [Figure 1] 1 illustrates a system for delivering a payload to a cell, according to some embodiments. [Figure 2A]1A-1D show various views of a device including a cartridge and a container for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 2B] 1A-1D show various views of a device including a cartridge and a container for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 2C] 1A-1D show various views of a device including a cartridge and a container for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 2D] 1A-1D show various views of a device including a cartridge and a container for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 2E] 1A-1D show various views of a device including a cartridge and a container for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 3A] FIG. 1 shows a diagram of a cartridge for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 3B] FIG. 1 shows a diagram of a cartridge for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 4A] FIG. 1 shows a diagram of a chip and gasket assembly for use with a cartridge for use in a system for delivering a payload to a cell, according to some embodiments. [Figure 4B] FIG. 1 shows a diagram of a chip and gasket assembly for use with a cartridge for use in a system for delivering a payload to a cell, according to some embodiments. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0028] Described below are exemplary embodiments of cartridges for use in partially or fully automated intracellular payload delivery systems, as well as related devices, systems, methods and techniques.

[0029] In the following description, example systems, methods, techniques, parameters, etc. are set forth. However, it should be recognized that such description is not intended to limit the scope of the present disclosure, but is provided as a description of example embodiments.

[0030] definition For purposes of interpreting this specification, the following definitions shall apply and whenever appropriate, terms used in the singular shall include the plural and vice versa. In the event that any definition set forth below conflicts with any document incorporated herein by reference, the definition set forth below shall control.

[0031] The term "pore" as used herein refers to an opening, including but not limited to a hole, crevice, cavity, aperture, crevice, gap, or perforation in a material. In some examples, the term refers to a hole in a surface of the present disclosure (where indicated and / or where apparent to one of skill in the art in light of the present disclosure). In other examples, the pore may refer to a hole in a cell membrane (where indicated and / or where apparent to one of skill in the art in light of the present disclosure).

[0032] The term "filter" as used herein refers to a porous body that selectively allows passage of fluid through the pores. In some embodiments, the term refers to a surface or membrane that contains pores.

[0033] In the description herein, various elements are described using terms such as first, second, etc., but these elements should not be limited to such terms. These terms are used only to distinguish one element from another.

[0034] For any of the structural and functional properties described herein, methods for determining these properties are known in the art.

[0035] All references cited herein, including patent applications and publications, are incorporated by reference in their entirety.

[0036] Cartridges for use in intracellular payload delivery systems FIG. 1 shows a schematic diagram of an intracellular payload delivery system 100.

[0037] In some embodiments, system 100 may share any one or more characteristics with any one of the systems described in WO2019 / 126212 and / or WO2020 / 210162, the entire contents of each of which are incorporated herein by reference. In short, an intracellular payload delivery system may be capable of delivering a payload into a cell by forcing the cell through a constriction, such as a narrow microfluidic channel or narrow pore, thereby perturbing the membrane of the cell and allowing the payload to enter the cell.

[0038] WO2020 / 210162 describes a system for intracellular payload delivery that relies on a cartridge configured to receive a flow of cell suspension from a preparation in fluid communication with a flexible tube and configured to direct the flow of cell suspension to a drainage bag. As described herein, the cartridges disclosed herein may differ from those described in WO2020 / 210162 in that the cartridges described herein may be provided in the form of a cap device configured to connect to a container, such as a test tube, and may include an input reservoir formed as an integral part of the cap device.

[0039] In some embodiments of system 100, a constriction, such as a narrow microfluidic channel or narrow hole, may be provided in a microfluidic chip or filter (which may be referred to as a constriction-containing element), which may be attached in fluid communication to system 100. In some embodiments, a microfluidic chip or filter (or any other element containing one or more constrictions configured to perturb a membrane of a cell) may be provided and fluidly coupled to a system such as system 100 by a cartridge. A cartridge may be any device configured to house a constriction-containing element, such as a microfluidic chip or filter, and / or configured to facilitate fluidic coupling of the element (e.g., chip or filter) to another portion of an intracellular payload delivery system, such as system 100.

[0040] 1, the system 100 may include a cartridge 300, a container 204, a cell suspension source 108, and a pressure source 110. The cartridge 300 and the container 204 together may form the device 200.

[0041] Cartridge 300 may be formed as a cap for container 204 and may be configured to be attached to container 204 by a threaded connection, a crimp connection, a snap-fit ​​connection, or any other suitable attachment means. When cartridge 300 is in an attached position relative to container 204, it may cover an opening in container 204, which in the arrangement shown in FIG. 1 may be an opening at the top of container 204.

[0042] The container 204 may be any suitable container having one or more exterior walls surrounding an interior cavity and having at least one opening to the interior cavity, the opening configured to mate with the cartridge 300. In some embodiments, the container 204 may be a tube, such as a test tube, a culture tube, a sample tube, or a centrifuge tube. In some embodiments, the container 204 may be a microcentrifuge tube.

[0043] The cartridge 300 may include both an input reservoir and a storage portion. The input reservoir may include a cavity formed in the cartridge that is configured to contain the cell suspension before the cell suspension is flowed through a constriction-containing element, such as a microfluidic chip or a filter. The storage portion may include a portion of the cartridge 300 that is configured to receive and contain the constriction-containing element itself. The input reservoir and storage portion may be fluidly connected to one another, for example, by one or more internal channels, such that the cell suspension may flow from the input reservoir to the storage portion and into the constriction-containing element contained in the storage portion. After flowing through the constriction-containing element, the cell suspension may flow out of the constriction-containing element and back into the storage portion. The cell suspension may then flow from the storage portion out of the cartridge and into the cavity of the vessel, for example, by one or more internal channels.

[0044] System 100 may further include a cell suspension source 108, which may include any source (e.g., a reservoir, tank, vessel, etc.) that may deliver a cell suspension to cartridge 300. In some embodiments, an input opening to an input reservoir of cartridge 300 may be configured to be fluidly coupled (e.g., by being selectively connectable and disconnectable) to cell suspension source 108 such that the cell suspension may flow from cell suspension source 108 to the input reservoir of cartridge 300.

[0045] System 100 may further include pressure source 110, which may include any pressure source configured to apply pressure to the cell suspension in the input reservoir of cartridge 300. In some embodiments, pressure source 110 may be provided as part of the same fluid source mechanism as cell suspension source 108. In some embodiments, pressure source 110 may be configured to be coupled (e.g., by being selectively connectable and disconnectable) to the input reservoir of cartridge 300 to apply pressurized gas, pressurized liquid, and / or mechanical pressure to the cell suspension in the input reservoir of cartridge 300. Application of pressure to the cell suspension in the input reservoir of cartridge 300 may cause the cell suspension to flow out of the input reservoir and through one or more constrictions of a constriction-containing element contained in a housing portion of cartridge 300. In some embodiments, pressure source 110 may share any one or more characteristics in common with the pressure control module and / or other pressure control components described in WO2019 / 126212. As noted above, WO2019 / 126212 is incorporated by reference herein in its entirety and for all purposes.

[0046] Described below are exemplary embodiments of improved cartridges for use in a system for intracellular payload delivery, such as cartridge 300 for use in system 100.

[0047] 2A-2E show various views of a device 200 including a cartridge 300 and a container 204 for use in a system 100 for delivering a payload to a cell, according to some embodiments.

[0048] The cartridge 300 may be configured to house one or more constriction-containing elements, such as a constriction filter (containing one or more constriction holes) or a constriction microfluidic chip (containing one or more constriction microfluidic channels). (A constriction filter according to some embodiments is disclosed in Application No. WO / 2017 / 041050A1, which is incorporated by reference in its entirety.) Note that in some embodiments, a constriction microfluidic channel or constriction hole may simply be referred to as a "constriction" or a "cell deformation constriction." A constriction-containing element may be any component, device, or part, etc., that has a channel, passage, or other opening (e.g., a constriction) that is smaller in diameter than the cells of the cell suspension passing through the element, such that when pressure is applied to force the cells through the opening, the opening constricts the cells, causing a perturbation in the cell's membrane. In some embodiments, device 200 and / or cartridge 300 may include an integral constriction filter or an integral constriction microfluidic channel configured to constrict cells, although in some embodiments cartridge 200 may be configured to house a separate element (e.g., a chip or filter) that itself has a constriction hole or constriction fluidic channel. In either case, device 200 and cartridge 300 may define a portion of a flow path of a system for delivering a payload to cells, such as system 100, whereby a cell suspension may flow into cartridge 300, which may then flow out through cartridge 300 and into reservoir 204 (and / or into any other suitable downstream flow path component).

[0049] FIG. 2A shows a perspective view of device 200. FIG. 2B shows an exploded perspective view of device 200. FIG. 2C shows a first side view of device 200. FIG. 2D shows a second side view of device 200, rotated 90 degrees from the first side view shown in FIG. 2C. FIG. 2E shows a top view of device 200. In each of the views of FIG. 2A-E, device 200 is shown with a chip and gasket assembly inserted into housing portion 306 of cartridge 300.

[0050] 2A-2C, portions of the cartridge 300 (e.g., cartridge body 302) and reservoir 202 are shown as partially transparent. In some embodiments, some or all portions of the cartridge 300 and / or reservoir 202 may be transparent, translucent, or opaque.

[0051] 2A-2E, the vessel 204 takes the form of a tube, such as a test tube, culture tube, sample tube, or centrifuge tube. In some embodiments, the vessel 204 may be a microcentrifuge tube. As such, the vessel 204 may be a prefabricated component with additional configuration for housing a chip.

[0052] In some embodiments, the container 204 may include one or more components made of metal, plastic, polymer, and / or glass, hi some embodiments, the container 204 may include one or more components made of polycarbonate, polypropylene, and / or polymethylmethacrylate.

[0053] In some embodiments, the interior cavity of the container 204 may have a volume of 1 mL or more, 1.5 mL or more, 2 mL or more, 2.5 mL or more, 3 mL or more, or 3.5 mL or more. In some embodiments, the interior cavity of the container 204 may have a volume of 1 mL or less, 1.5 mL or less, 2 mL or less, 2.5 mL or less, 3 mL or less, or 3.5 mL or less.

[0054] As further shown in Figures 2A-2E, cartridge 300 (shown and described in further detail with reference to Figures 3A-3B) may be configured to mate with opening 205 of container 204, for example, by taking the form of a cap for container 204. In some embodiments, cartridge 300 may be configured to mate with opening 205 of container 204 by a crimp connection, a snap connection, a twist lock connection, or any other suitable connection. In some embodiments, cartridge 300 may be configured to be held against opening 205 of container 204 by one or more tube manipulation devices, such as a robotic arm, during processing of a cell suspension through cartridge 300.

[0055] Unlike cartridge systems that may be configured to be attached to an input reservoir by a flexible tube or other fluidics components, as described above, the cartridges disclosed herein may include an input reservoir that is physically integrated with the cartridge itself. The input reservoir may be a reservoir configured to contain the cell suspension before it flows through the constriction-containing element. The cartridges disclosed herein may be provided in the form of a cap device that is configured to couple to and cover the opening of a container, such as a test tube, culture tube, sample tube, or centrifuge tube. The use of an integrated input reservoir facilitates integration with existing laboratory devices, such as readily available pressure sources. Furthermore, the timing and volume reduction potential of using the embodiments described herein may be particularly useful, for example, in research use cases where volumes are small and consumables can be used quickly with multiple small-volume experiments. The container (e.g., tube) to which the cartridge may be attached as a cap may serve as an output reservoir to contain the cell suspension after processing through the constriction-containing element. In some embodiments, when the cartridge is in an attached position with a vessel (e.g., tube), a portion of the cartridge may be disposed inside the interior cavity of the vessel, for example by projecting downwardly into the interior of a test tube. This configuration may facilitate retrieval of the tubular vessel and further facilitate its use in a laboratory setting.

[0056] Thus, when the cartridge is attached to the open end of a vessel, such as a laboratory tube, the device formed by the combination of the cartridge and vessel may include an input reservoir, a housing portion configured to receive and hold a constriction-containing element, and an output reservoir provided in the form of a vessel, such as a laboratory tube. All three components may be in fluid communication with each other, allowing the cell suspension to flow from the input reservoir to the housing portion (and into and out of the constriction-containing element contained in the housing portion) and then from the housing portion to the vessel (e.g., tube). After the cell suspension has flowed into the vessel (e.g., tube), the cartridge may be removed from the opening of the vessel and the vessel (containing the processed cell suspension) may be used for further processing of the cell suspension, may be capped, and / or may be stored in any suitable manner. If the vessel is provided in the form of a standard laboratory tube, the tube may be easily used for downstream processing with one or more standard-compatible additional devices, caps, and / or storage devices.

[0057] 2A and 2C-2E, the cartridge 300 can be configured such that when assembled with the container 204 by being placed in the assembly position, the cartridge body 302 extends downwardly into the internal cavity of the container 204. In some embodiments, a width of a portion of the cartridge body 302 can be less than an inner diameter of the internal cavity of the container 204 such that the portion of the cartridge body 302 can extend downwardly into the internal cavity of the container 204. In some embodiments, the cartridge body 302 can include a rim portion that is wider than the portion that extends into the internal cavity of the container 204, such that the rim portion can be located outside of the opening 205 of the container 204 (e.g., can be located above the opening 205) while at the same time the portion of the cartridge body 302 extends into the internal cavity of the container 204. In some embodiments, the cartridge 300, when in an assembled position with the container 204, may extend into the interior cavity of the container 204 by 30% or more, 40% or more, 50% or more, 60% or more, or 70% or more of the total height dimension of the interior cavity of the container 204 (total height dimension in the direction in which the cartridge 300 extends into the interior cavity). In some embodiments, the cartridge 300, when in an assembled position with the container 204, may extend into the interior cavity of the container 204 by 30% or less, 40% or less, 50% or less, 60% or less, or 70% or less of the total height dimension of the interior cavity of the container 204 (total height dimension in the direction in which the cartridge 300 extends into the interior cavity). In some embodiments, the cartridge 300, when in an assembled position with the container 204, may extend into the interior cavity of the container 204 by 0.5 cm or more, 1 cm or more, 1.5 cm or more, 2 cm or more, or 2.5 cm or more. In some embodiments, the cartridge 300, when in an assembled position with the container 204, may extend into the interior cavity of the container 204 by 0.5 cm or less, 1 cm or less, 1.5 cm or less, 2 cm or less, or 2.5 cm or less.

[0058] In some embodiments, when in an assembled position with the container 204, 50% or more, 60% or more, 70% or more, 80% or more, 90% or more, 95% or more, or 99% or more of the height of the cartridge 300 may extend into the interior cavity of the container 204. In some embodiments, when in an assembled position with the container 204, 50% or less, 60% or less, 70% or less, 80% or less, 90% or less, 95% or less, or 99% or less of the height of the cartridge 300 may extend into the interior cavity of the container 204.

[0059] 2B , in some embodiments, the cartridge 300 may comprise a cartridge body 302 that includes an input reservoir 304 and a storage portion 306. The input reservoir 304 may be configured to receive and hold the cell suspension before the cell suspension flows from the input reservoir 304 to the storage portion 306.

[0060] In some embodiments, the input reservoir may have a tapered shape such that under gravity and / or pressure (e.g., liquid pressure, gas pressure, and / or mechanical pressure), fluid may flow out of the bottom of the input reservoir 304 and into one or more channels. In some embodiments, the input reservoir 306 may have an internal volume of 1 mL or less, 1.5 mL or less, 2 mL or less, 2.5 mL or less, or 3 mL or less. In some embodiments, the input reservoir 306 may have an internal volume of 1 mL or more, 1.5 mL or more, 2 mL or more, 2.5 mL or more, or 3 mL or more.

[0061] The storage portion 306 may be configured to receive and hold a constriction-containing element, such as a tip and gasket assembly 400 (shown and described in further detail with reference to FIGS. 4A-4B ), and to allow a cell suspension to flow from the input reservoir 304 into the storage portion and through one or more channels in the storage portion into the constriction-containing element. After the cell suspension flows through the constriction-containing element, the suspension may flow back into the one or more channels in the storage portion. The one or more channels in the storage portion may be open to the exterior of the cartridge 300, allowing the cell suspension to flow out of the cartridge 300 and into the interior cavity of the vessel 204.

[0062] Cartridge 300 may include a housing cover 308 that may be configured to cover housing portion 306 and to hold the constriction-containing element in place within housing portion 306 .

[0063] The storage portion cover 308 may be an element configured to be positioned alongside the one or more stenosis-containing elements, to press the one or more stenosis-containing elements against the storage portion 306, and / or to otherwise hold the one or more stenosis-containing elements in place. In some embodiments, the storage portion cover 308 may be configured to apply an inward force to the one or more stenosis-containing elements, pressing them against the storage portion 306, by one or more springs or other compressible or deformable components, such as one or more rubber O-rings or gaskets. In some embodiments, the storage portion cover 308 may be configured to directly press against a surface of one or more of the stenosis-containing elements. In some embodiments, the storage portion cover 308 may help ensure that the pressure of the fluid forced through the one or more stenosis-containing elements does not cause delamination of the one or more stenosis-containing elements, and delamination of the stenosis-containing elements may be prevented by pressing the storage portion cover 308 against one side of the stenosis-containing elements with force.

[0064] For example, the storage portion cover 308 may be attachable and detachable to the storage portion 306 by a sliding connection, a threaded connection, a hinge connection, a tab-slot connection, a locking mechanism, by one or more screws, by one or more cams, or in any other suitable manner so that the storage portion cover 308 can be removed to replace the gasket 404 and / or the constriction-containing element 402.

[0065] 2A-2E, a sliding connection such as that shown in FIG. 2A-2E may allow the storage portion cover 308 to slide laterally (in the direction of the double arrow shown in FIG. 2B) along the storage portion 306 such that a lip of the storage portion cover 308 may slidingly fit into a corresponding lip, protrusion, groove, or tooth of the storage portion 306. In some embodiments, a removable cover configured to slide onto and off of the body of the storage portion may be configured to completely surround the storage portion, which in some embodiments avoids the need for mating grooves, teeth, or the like.

[0066] Such a sliding connection may be removed with minimal lateral force (e.g., in the sliding direction), but may provide significant strength in a direction perpendicular to the sliding direction, extending away from the housing portion in which the cover is located. Thus, a cover having a sliding connection may provide superior pressure durability over other connection mechanisms that may be used to manually assemble the cartridge, such as attaching a cover with a threaded component, even though the cover may be easily removed by hand. In some embodiments, in addition to or in the alternative to one or more removable covers, the cartridge may be configured to securely house one or more stenosis-containing elements without the use of a removable cover.

[0067] In some embodiments, the storage portion cover 308 may be configured to be slidably removed from the storage portion 306, but may be configured to be unable to slide out of the storage portion 306 when the cartridge 300 is in an assembled position with the receptacle 204. As shown in FIGS. 2A-2E, when the cartridge 300 is assembled with the receptacle 204 such that the cartridge body 302 extends downwardly into the interior cavity of the receptacle 204, the walls of the receptacle 204 may block the storage portion cover 308 such that it cannot slide out of either side of the storage portion 306. This arrangement may help ensure that the storage portion cover 308 is not accidentally removed from the storage portion during operation.

[0068] 2B, cartridge 300 can be configured to allow cell suspension fluid to enter (e.g., flow into) input reservoir 304 through an opening, such as top opening 305. In some embodiments, input reservoir 304 can have an open top end such that the cell suspension can flow into input reservoir 304 from the top.

[0069] In some embodiments, the cartridge 300 may include a lid 326 that may cover the opening of the input reservoir 304. The lid 326 may be configured to mate with the opening of the input reservoir 304 via a crimp connection, a snap connection, a twist lock connection, or any other suitable connection. The lid 326 may be a bendable lid including an upper portion 327 and a lower portion 329, where the upper portion 327 may be bent up to an open position that allows access to the input reservoir 304 and then bent down to a closed position that confines the cell suspension within the input reservoir 304. In some embodiments, the lid 326 may be placed in the closed position before pressure is applied to the cell suspension in the input reservoir 304. In some embodiments, the lid 326 may include a filter 324, which may be configured to prevent liquid (e.g., the cell suspension) from flowing back from the cartridge toward the cell suspension source and / or the pressure source. Air pressure may be provided above the filter 324, for example, from a pressure source.

[0070] 2B and 2C, cartridge 300 may include two or more O-rings, including O-ring 320 and O-ring 322. O-ring 320 may be disposed between lid 326 and cartridge body 302 to form a seal therebetween. O-ring 322 may be disposed on the opposite side of cartridge body 302 from lid 326 to form a seal between lid 326 and another component of system 100, such as cell suspension source 108 and / or pressure source 110. In some embodiments, O-ring 322 may be pressed against another component of system 100 (e.g., by a robotic arm) to form a seal and allow positive pressure to be applied to the cell suspension in input reservoir 304.

[0071] 3A-3B show diagrams of a cartridge 300 for use in the system 100 for delivering a payload to a cell, according to some embodiments.

[0072] Figure 3A shows a side view of cartridge 300. Figure 3B shows an enlarged side view of the lower portion of device 300, focusing on housing portion 306 and housing portion cover 308. In both Figures 3A and 3B, cartridge 300 is shown with a chip and gasket assembly inserted therein.

[0073] 3A and 3B, portions of the cartridge 300 (e.g., the cartridge body 302) are shown as partially transparent. In some embodiments, some or all portions of the cartridge 300 can be transparent, translucent, or opaque.

[0074] 3A and 3B provide illustrations of the internal channels formed within the cartridge body 302 that facilitate the flow of the cell suspension through the cartridge 300.

[0075] As shown, the input channel 310 fluidly connects the input reservoir 304 to the storage portion 306. In the configuration shown, the input channel 310 has a first vertical portion and then a second horizontal portion, beginning at the outlet of the input reservoir 304 and terminating in a portion configured to hold a gasket of a constriction-containing element in the storage portion 306 (discussed in more detail with respect to FIGS. 4A-4B). In some embodiments, all or a portion of the input channel 310 may have a diameter of 0.05 mm or less, 0.1 mm or less, 0.5 mm or less, 1 mm or less, 1.5 mm or less, 2 mm or less, or 2.5 mm or less. In some embodiments, all or a portion of the input channel 310 may have a diameter of 0.05 mm or more, 0.1 mm or more, 0.5 mm or more, 1 mm or more, 1.5 mm or more, 2 mm or more, or 2.5 mm or more.

[0076] As shown, the outlet channel 312 begins at a portion configured to hold the gasket of the constriction-containing element in the receiving portion 306 and terminates at an opening to the exterior of the cartridge 300. Thus, the cell suspension that has passed through the constriction-containing element may flow through the outlet channel 312 and exit the cartridge 300, for example, dripping downwards by gravity, to be collected in the container (e.g., container 204) into which the cartridge 300 is inserted. In some embodiments, all or a portion of the outlet channel 312 may have a diameter of 0.05 mm or less, 0.1 mm or less, 0.5 mm or less, 1 mm or less, 1.5 mm or less, 2 mm or less, or 2.5 mm or less. In some embodiments, all or a portion of the outlet channel 312 may have a diameter of 0.05 mm or more, 0.1 mm or more, 0.5 mm or more, 1 mm or more, 1.5 mm or more, 2 mm or more, or 2.5 mm or more.

[0077] In some embodiments, the height of cartridge 300 (e.g., the vertical height as shown in FIG. 2A) can be 0.5 mm or less, 0.6 mm or less, 0.75 mm or less, 1 mm or less, 5 mm or less, 1 cm or less, 5 cm or less, or 10 cm or less. In some embodiments, the height of cartridge 300 can be 0.5 mm or more, 0.6 mm or more, 0.75 mm or more, 1 mm or more, 5 mm or more, 1 cm or more, 5 cm or more, or 10 cm or more.

[0078] In some embodiments, when the cartridge 300 is in an assembled position relative to the container 204, the width of the cartridge 300 (e.g., the width in a direction extending horizontally in FIG. 2A ) can be less than or equal to a width measured at a portion of the cartridge 300 that extends into the interior cavity of the container 204. In some embodiments, when the cartridge 300 is in an assembled position relative to the container 204, the width of the cartridge 300 can be less than or equal to a width measured at a portion of the cartridge 300 that extends into the interior cavity of the container 204.

[0079] 4A-4B show diagrams of a chip and gasket assembly 400 for use with a cartridge for use in a system 100 for delivering a payload to a cell, according to some embodiments. The chip and gasket assembly 400 can be the same as the chip and gasket assemblies shown in FIGS. 2A-2D, 3A, and 3B. Although FIGS. 4A-4B show an exemplary microfluidic chip including multiple constrictions configured to induce cell membrane perturbations, it will be understood in light of the disclosure herein that a microfluidic filter configured to induce cell membrane perturbations may additionally or alternatively be used.

[0080] Figure 4A shows a perspective view of chip and gasket assembly 400. Figure 4B shows a top view of chip and gasket assembly 400 with gasket 404 on the foreground.

[0081] 3A and 3B, portions of the cartridge 300 (e.g., the cartridge body 302) are shown as partially transparent. In some embodiments, some or all portions of the cartridge 300 can be transparent, translucent, or opaque.

[0082] 4A, the chip and gasket assembly 400 can include a chip 402 and a gasket 404, which can be disposed adjacent to one another in a stacked configuration. The chip 402 itself can include a constriction layer 402a and a cover layer 402b.

[0083] Constriction layer 402a may be a layer in which one or more cell deformation constrictions are formed, for example, by etching or machining into the layer. Constriction layer 402a may be formed of silicon in some embodiments.

[0084] The cover layer 402b may be disposed against the constriction layer 402a to cover one or more cavities and / or constrictions formed in the constriction layer 402a. Thus, the cover layer 402b may form a "lid" on top of the constriction layer 402a to seal the volume through which the cell suspension flows through the constriction layer 402a. As shown in FIGS. 4A and 4B, the cover layer 402a may include a chip inlet channel 408 and a chip outlet channel 410, each formed as a through-hole in the cover layer 402a to allow the cell suspension to flow through the channel. The cell suspension may flow into the chip 402, through the chip inlet channel 408 into the constriction layer 402a, out of the constriction layer 402a, into the chip outlet channel 410, and out of the chip 402. The cover layer 402b may be formed of glass in some embodiments. The cover layer 402b may be bonded to the constriction layer 402a by any suitable means, such as with an adhesive.

[0085] In some embodiments, one or both of the chip inlet channel 408 and the chip outlet channel 410 can have a diameter of 0.2 mm or less, 0.4 mm or less, 0.6 mm or less, 0.8 mm or less, 1 mm or less, or 1.2 mm or less. In some embodiments, one or both of the chip inlet channel 408 and the chip outlet channel 410 can have a diameter of 0.2 mm or more, 0.4 mm or more, 0.6 mm or more, 0.8 mm or more, 1 mm or more, or 1.2 mm or more.

[0086] The gasket 404 can be configured to be disposed against the cover layer 402a. The gasket 404 can provide a channel fluidly connecting the tip 402 to a channel of the housing portion 306 and form a seal against the housing portion 306 to allow fluid to flow from the housing portion 306 to the tip 402 and from the tip 402 back to the housing portion 306.

[0087] 4A and 4B, the gasket 404 can include a gasket inlet channel 406 and a gasket outlet channel 412, each of which can be formed as a through-hole in the gasket 404 to allow the cell suspension to flow therethrough. The cell suspension can flow into the chip 402 through the gasket inlet channel 406 and out of the chip 402 into and through the gasket outlet channel 412.

[0088] In some embodiments, one or both of the gasket inlet channel 406 and the gasket outlet channel 412 can have a cross-sectional dimension (eg, hydraulic diameter) between 0.1 mm and 2.0 mm.

[0089] The gasket 404, in some embodiments, may be formed from one or more of silicone, rubber, and / or thermoplastic elastomers.

[0090] Thus, the cell suspension may flow from input reservoir 304 to input channel 310, from input channel 310 to gasket inlet channel 406, from gasket inlet channel 406 to chip inlet channel 408, from chip inlet channel 408 to constriction layer 402a, from constriction layer 402a to chip outlet channel 410, from chip outlet channel 410 to gasket outlet channel 412, from gasket outlet channel 412 to exhaust channel 312, and from exhaust channel 312 out of cartridge 300 and into the interior cavity of vessel 204. Buffers or other fluids may also flow along the same or similar flow paths.

[0091] In some embodiments, in addition to or as an alternative to using a gasket such as gasket 414, a seal for the fluid connection between the cartridge and the constriction-containing element may be created using other sealing options, for example, an O-ring, overmolding, chemical bonding, and / or a mechanical interlock may be used.

[0092] In some embodiments, the tip 402 may have a length and / or width of 50 mm or less, 25 mm or less, 10 mm or less, 5 mm or less, 4 mm or less, 3 mm or less, 2 mm or less, or 1 mm or less. In some embodiments, the tip 402 may have a length and / or width of 50 mm or more, 25 mm or more, 10 mm or more, 5 mm or more, 4 mm or more, 3 mm or more, 2 mm or more, or 1 mm or more. In some embodiments, the tip 402 may have a footprint of 4.1 mm by 4.1 mm.

[0093] In some embodiments, the tip 402 may have a thickness of 0.1 mm or less, 0.5 mm or less, 1 mm or less, 2 mm or less, 3 mm or less, or 5 mm or less. In some embodiments, the tip 402 may have a thickness of 0.1 mm or less, 0.5 mm or less, 1 mm or less, 2 mm or less, 3 mm or less, or 5 mm or less.

[0094] In some embodiments, the constriction layer 402a may have a thickness of 0.2 mm or less, 0.4 mm or less, 0.6 mm or less, 0.8 mm or less, 1 mm or less, 1.5 mm or less, or 2 mm or less. In some embodiments, the constriction layer 402a may have a thickness of 0.2 mm or less, 0.4 mm or less, 0.6 mm or less, 0.8 mm or less, 1 mm or less, 1.5 mm or less, or 2 mm or less.

[0095] In some embodiments, the cover layer 402b may have a thickness of 0.2 mm or less, 0.4 mm or less, 0.6 mm or less, 0.8 mm or less, 1 mm or less, 1.5 mm or less, or 2 mm or less. In some embodiments, the cover layer 402b may have a thickness of 0.2 mm or less, 0.4 mm or less, 0.6 mm or less, 0.8 mm or less, 1 mm or less, 1.5 mm or less, or 2 mm or less.

[0096] In some embodiments, the width of the constriction (e.g., constricted channel or constricted hole) of the constriction-containing element (e.g., chip 402) can be 0.25 μm or less, 0.5 μm or less, 1 μm or less, 5 μm or less, 10 μm or less, 20 μm or less, or 50 μm or less. In some embodiments, the width of the constriction (e.g., constricted channel or constricted hole) of the constriction-containing element (e.g., chip 402) can be 0.25 μm or more, 0.5 μm or more, 1 μm or more, 5 μm or more, 10 μm or more, 20 μm or more, or 50 μm or more.

[0097] In some embodiments, the length of the constriction (e.g., constriction channel or constriction hole) of the constriction-containing element (e.g., chip 402) can be 1 μm or less, 5 μm or less, 10 μm or less, 20 μm or less, 30 μm or less, 40 μm or less, 50 μm or less, or 100 μm or less. In some embodiments, the length of the constriction (e.g., constriction channel or constriction hole) of the constriction-containing element (e.g., chip 402) can be 1 μm or more, 5 μm or more, 10 μm or more, 20 μm or more, 30 μm or more, 40 μm or more, 50 μm or more, or 100 μm or more.

[0098] In some embodiments, the depth of the constriction (e.g., constriction channel or constriction hole) of the constriction-containing element (e.g., chip 402) can be 10 μm or less, 15 μm or less, 20 μm or less, 50 μm or less, 80 μm or less, 100 μm or less, or 200 μm or less. In some embodiments, the depth of the constriction (e.g., constriction channel or constriction hole) of the constriction-containing element (e.g., chip 402) can be 10 μm or more, 15 μm or more, 20 μm or more, 50 μm or more, 80 μm or more, 100 μm or more, or 200 μm or more.

[0099] In some embodiments, the gasket 404 can have a length and / or width of 50 mm or less, 25 mm or less, 10 mm or less, 5 mm or less, 4 mm or less, 3 mm or less, 2 mm or less, or 1 mm or less. In some embodiments, the gasket 404 can have a length and / or width of 50 mm or more, 25 mm or more, 10 mm or more, 5 mm or more, 4 mm or more, 3 mm or more, 2 mm or more, or 1 mm or more. In some embodiments, the gasket 404 can have a footprint of 4.1 mm by 4.1 mm.

[0100] In some embodiments, the gasket 404 can have a thickness of 0.25 mm or less, 0.5 mm or less, 0.75 mm or less, 1 mm or less, 1.25 mm or less, 1.5 mm or less, 1.75 mm or less, 2 mm or less, or 2.25 mm or less. In some embodiments, the gasket 404 can have a thickness of 0.25 mm or less, 0.5 mm or less, 0.75 mm or less, 1 mm or less, 1.25 mm or less, 1.5 mm or less, 1.75 mm or less, 2 mm or less, or 2.25 mm or less.

[0101] In some embodiments, multiple constriction-containing elements within the cartridge 300 may be arranged in series, such that the flow path through the cartridge 300 may be a single linear path. In some embodiments, multiple constriction-containing elements within the cartridge 300 may be arranged in parallel, such that the flow path within the cartridge 300 may diverge into multiple parallel segments as the fluid travels through the cartridge 300 and then reconverge before exiting the cartridge 300. In some embodiments, three or more constriction-containing elements may be arranged within the cartridge 300, with one or more of the elements being arranged in series with another one of the constriction-containing elements and one or more of the elements being arranged in parallel with another one of the constriction-containing elements.

[0102] In some embodiments, the cartridge 300 may be configured to house blank placeholder elements in place of functional constriction-containing elements, which may not include any channels or holes or may be otherwise configured to be unable to flow through the portion of the cartridge 300 that houses the placeholder elements. The use of blank placeholder elements allows the cartridge 300 to effect fluid flow through a reduced number of constriction-containing elements at a time, or through only one constriction-containing element at a time, thereby eliminating the need to use the system with a maximum capacity of constriction-containing elements all the time.

[0103] In some embodiments, device 200 may be configured to force fluid through cartridge 300 at pressures of 1 PSI or less, 5 PSI or less, 10 PSI or less, 25 PSI or less, 50 PSI or less, 75 PSI or less, 100 PSI or less, 125 PSI or less, 150 PSI or less, or 200 PSI or less. In some embodiments, device 200 may be configured to force fluid through cartridge 300 at pressures of 1 PSI or more, 5 PSI or more, 10 PSI or more, 25 PSI or more, 50 PSI or more, 75 PSI or more, 100 PSI or more, 125 PSI or more, 150 PSI or more, or 200 PSI or more. In some embodiments, device 200 may be configured for use with a constriction-containing element (e.g., a constriction-containing element) that can individually (e.g., on a "per chip" basis) provide a throughput of 50 mL or less, 100 mL or less, 150 mL or less, 200 mL or less, 250 mL or less, 300 mL or less, or 400 mL or less of red blood cell suspension per minute. In some embodiments, device 200 may be configured for use with a constriction-containing element (e.g., a constriction-containing element) that can individually (e.g., on a "per chip" basis) provide a throughput of 50 mL or more, 100 mL or more, 150 mL or more, 200 mL or more, 250 mL or more, 300 mL or more, or 400 mL or more of red blood cell suspension per minute.

[0104] In some embodiments, device 200 may be configured for use with a constriction-containing element (e.g., a constriction-containing element) that can provide a throughput of peripheral blood mononuclear cell suspension of 25 mL or less, 50 mL or less, 75 mL or less, 100 mL or less, 125 mL or less, 150 mL or less, or 200 mL or less, individually (e.g., on a "per tip" basis). In some embodiments, device 200 may be configured for use with a constriction-containing element (e.g., a constriction-containing element) that can provide a throughput of peripheral blood mononuclear cell suspension of 25 mL or more, 50 mL or more, 75 mL or more, 100 mL or more, 125 mL or more, 150 mL or more, or 200 mL or more, individually (e.g., on a "per tip" basis).

[0105] In some embodiments, device 200 may have a total fluid throughput of 0.01 L / min or less, 0.05 L / min or less, 0.1 L / min or less, 0.25 L / min or less, 0.5 L / min or less, or 1 L / min or less (e.g., including all constriction-containing elements contained in cartridge 300). In some embodiments, device 200 may have a total fluid throughput of 0.01 L / min or more, 0.05 L / min or more, 0.1 L / min or more, 0.25 L / min or more, 0.5 L / min or more, or 1 L / min or more (e.g., including all constriction-containing elements contained in cartridge 300).

Claims

1. 1. A device for facilitating delivery of a payload to cells of a cell suspension, the device comprising: a container including an interior cavity and an opening; A cartridge; the cartridge is configured to couple to the opening of the container when in an assembled position relative to the container, the cartridge comprising: an input reservoir configured to receive a cell suspension; a housing portion configured to house a constriction-containing element; the constriction-containing element comprises a constriction configured to perturb membranes of cells of the cell suspension to facilitate delivery of a payload to the cells; the containment portion is fluidly connected to the input reservoir, thereby allowing the cell suspension to flow from the input reservoir to the containment portion; the storage portion is directly fluidly connected to a cartridge outlet, thereby allowing the cell suspension to flow from the storage portion into the interior cavity of the vessel; The device.

2. The device of claim 1 , wherein a portion of the storage portion of the cartridge is disposed inside the internal cavity of the container when the cartridge is in the assembled position relative to the container.

3. The device of claim 1 , wherein a portion of the input reservoir of the cartridge is disposed inside the interior cavity of the container when the cartridge is in the assembled position relative to the container.

4. The device of claim 1 , further comprising a cover, the cover being releasably attached to the storage portion of the cartridge and configured to hold the constriction-containing element in place when the constriction-containing element is stored in the storage portion.

5. The device of claim 4 , wherein the cover is configured to be slidably attached to and removed from the housing portion of the cartridge.

6. The device of claim 5 , wherein the cover is configured to slide in a direction perpendicular to the direction in which the layers of the constriction-containing elements are stacked on top of each other.

7. The device of claim 4 , wherein the cover is configured to be prevented from removal from the storage portion by an inner wall of the container when the cartridge is in the assembled position relative to the container.

8. The device of claim 1 , wherein the cartridge comprises an input reservoir cover, the input reservoir cover being movable between an open position and a closed position to selectively expose and enclose an interior of the input reservoir.

9. 9. The device of claim 8, wherein the cartridge includes a first O-ring disposed against a surface of the input reservoir cover opposite the interior of the input reservoir when the input reservoir cover is in the closed position.

10. The device of claim 8 , wherein the cartridge includes a second O-ring disposed between the input reservoir cover and a body portion of the cartridge.

11. 10. The device of claim 8, wherein the input reservoir cover includes a filter configured to prevent backflow of the cell suspension.

12. The device of claim 1 , wherein the cartridge has a total fluid throughput of greater than 0.05 L / min.

13. 10. The device of claim 1, wherein the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure greater than 100 PSI.

14. 1. A device for facilitating delivery of a payload to cells of a cell suspension, the device comprising: a container including an interior cavity and an opening; A cartridge; the cartridge is configured to couple to the opening of the container when in an assembled position relative to the container, the cartridge comprising: an input reservoir configured to receive a cell suspension; The storage area and a constriction-containing element; the constriction-containing element comprises a constriction configured to perturb membranes of cells of the cell suspension to facilitate delivery of a payload to the cells; the storage portion of the cartridge is fluidly connected to the input reservoir and the constriction-containing element, such that the cell suspension can flow from the input reservoir through the storage portion into the constriction-containing element, and the cell suspension can flow from the constriction-containing element through the storage portion and into the internal cavity of the container; The device.

15. The device of claim 14 , further comprising a gasket disposed between the housing portion and the constriction-containing element.

16. 15. The device of claim 14, wherein the constriction-containing element comprises a microfluidic chip containing a plurality of constrictions that perturb the membrane of the cell.

17. 15. The device of claim 14, wherein the constriction-containing element has a footprint of less than 5 mm x 5 mm.

18. The device of claim 14 , wherein a portion of the storage portion of the cartridge is disposed inside the internal cavity of the container when the cartridge is in the assembled position relative to the container.

19. The device of claim 14 , wherein a portion of the input reservoir of the cartridge is disposed inside the internal cavity of the container when the cartridge is in the assembled position relative to the container.

20. 15. The device of claim 14, wherein the cartridge further comprises a cover, the cover being releasably attached to the storage portion of the cartridge and configured to hold the constriction-containing element in place when the constriction-containing element is stored in the storage portion.

21. The device of claim 20 , wherein the cover is configured to be slidably attached to and removed from the housing portion of the cartridge.

22. 22. The device of claim 21, wherein the cover is configured to slide in a direction perpendicular to the direction in which the layers of the constriction-containing elements are stacked on top of each other.

23. The device of claim 20 , wherein the cover is configured to be prevented from removal from the storage portion by an inner wall of the container when the cartridge is in the assembled position relative to the container.

24. 15. The device of claim 14, wherein the cartridge comprises an input reservoir cover, the input reservoir cover being movable between an open position and a closed position to selectively expose and enclose an interior of the input reservoir.

25. 25. The device of claim 24, wherein the cartridge includes a first O-ring disposed against a surface of the input reservoir cover opposite the interior of the input reservoir when the input reservoir cover is in the closed position.

26. 25. The device of claim 24, wherein the cartridge includes a second O-ring disposed between the input reservoir cover and a body portion of the cartridge.

27. 25. The device of claim 24, wherein the input reservoir cover includes a filter configured to prevent backflow of the cell suspension.

28. The device of claim 14 , wherein the cartridge has a total fluid throughput of greater than 0.05 L / min.

29. 15. The device of claim 14, wherein the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure greater than 100 PSI.

30. 1. A cartridge for facilitating delivery of a payload to cells of a cell suspension, said cartridge comprising: an input reservoir configured to receive a cell suspension; a coupling portion configured to couple to an opening to an interior cavity of the container; a housing portion configured to house a constriction-containing element; the constriction-containing element comprises a constriction configured to perturb membranes of cells of the cell suspension to facilitate delivery of a payload to the cells; the containment portion is fluidly connected to the input reservoir, thereby allowing the cell suspension to flow from the input reservoir to the containment portion; the reservoir is fluidly connected to a cartridge outlet, thereby allowing the cell suspension to exit the cartridge; The cartridge.

31. The cartridge of claim 30, wherein a portion of the storage portion of the cartridge is disposed inside the internal cavity of the container when the cartridge is coupled to the container in an assembled position.

32. The cartridge of claim 30, wherein a portion of the input reservoir of the cartridge is disposed inside the internal cavity of the container when the cartridge is coupled to the container in an assembled position.

33. 31. The cartridge of claim 30, further comprising a cover, the cover being releasably attached to the storage portion of the cartridge and configured to hold the constriction-containing element in place when the constriction-containing element is stored in the storage portion.

34. 34. The cartridge of claim 33, wherein the cover is configured to be slidably attached to and removed from the storage portion of the cartridge.

35. 35. The cartridge of claim 34, wherein the cover is configured to slide in a direction perpendicular to the direction in which the layers of the constriction-containing elements are stacked on top of each other.

36. 34. The cartridge of claim 33, wherein when the cartridge is coupled to the container in an assembled position, the cover is configured to be prevented from removal from the storage portion by an inner wall of the container.

37. 31. The cartridge of claim 30, wherein the cartridge comprises a loading reservoir cover, the loading reservoir cover being movable between an open position and a closed position to selectively expose and enclose an interior of the loading reservoir.

38. 40. The cartridge of claim 37, wherein the cartridge includes a first O-ring disposed against a surface of the input reservoir cover opposite the interior of the input reservoir when the input reservoir cover is in the closed position.

39. 40. The cartridge of claim 37, wherein the cartridge includes a second O-ring disposed between the input reservoir cover and a body portion of the cartridge.

40. 40. The cartridge of claim 37, wherein the input reservoir cover comprises a filter configured to prevent backflow of the cell suspension.

41. 31. The cartridge of claim 30, wherein the cartridge has a total fluid throughput of greater than 0.05 L / min.

42. 31. The cartridge of claim 30, wherein the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure greater than 100 PSI.

43. The device of claim 12 , wherein the cartridge has a total fluid throughput of less than 0.5 L / min.

44. 30. The device of claim 28, wherein the cartridge has a total fluid throughput of less than 0.5 L / min.

45. 42. The cartridge of claim 41, wherein the cartridge has a total fluid throughput of less than 0.5 L / min.

46. 14. The device of claim 13, wherein the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure of less than 200 PSI.

47. 30. The device of claim 29, wherein the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure of less than 200 PSI.

48. 43. The cartridge of claim 42, wherein the cartridge is configured to allow the cell suspension to flow through the cartridge at a pressure of less than 200 PSI.

Images