Cartridge and device for use in a system for delivering a paload into cells

Abstract

A device for facilitating delivery of a payload to cells of a cell suspension includes a container and a cartridge configured to connect to an opening of the container when in an assembled position relative to the container. The cartridge includes an input reservoir configured to receive a cell suspension. The cartridge includes a receiving portion configured to receive a constriction containing element, where the constriction containing element includes a constriction configured to disrupt membranes of cells of the cell suspension to facilitate delivery of a payload to the cells, wherein the receiving portion is fluidly connected to the input reservoir to allow the cell suspension to flow from the input reservoir to the receiving portion, and wherein the receiving portion is fluidly connected to a cartridge outlet port to allow the cell suspension to flow from the receiving portion into the interior cavity of the container.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to and the benefit of U.S. Provisional Application No. 63 / 346,634, filed on May 27, 2022, the entire contents of which are hereby incorporated by reference in its entirety for all purposes. Technical Field

[0003] The present disclosure relates to systems for delivering payloads into cells, and more particularly to cassettes and devices for holding constriction containing elements having constriction channels or constriction pores for use in the systems to cause perturbations in the cell membrane to allow the payload to pass through the cell membrane. Background Art

[0004] In the ever-evolving field of cell therapy medicine, it is of great significance to controllably deliver various materials into cells. For example, various research and therapeutic applications may deliver peptides, nucleic acids, proteins, small molecules and nanomaterials through cell membranes and into cells. As discussed in WO2013059343, WO2015023982, PCT / US2015 / 058489, PCT / US2015 / 060689 and PCT / US2016 / 13113, contraction microfluidic channels can be used to deliver compounds and other payloads into cells. As disclosed in PCT / US18 / 66295, desktop laboratories and / or clinical systems can be configured to force cell suspensions to pass through boxes, wherein the boxes accommodate one or more contraction accommodating elements (e.g., parts, pieces, devices, components or the like, such as microfluidic chips or filters) with contraction channels or contraction holes to cause disturbances in the membranes of cells in cell suspensions. Summary of the invention

[0005] As explained above, some systems for intracellular payload delivery include a box configured to accommodate one or more contraction-containing elements (e.g., a microfluidic chip or a filter) having a contraction channel or contraction hole so as to cause disturbances in the membranes of cells in the cell suspension when the cell suspension flows through the box. However, these systems have boxes that can be improved. For example, such systems cannot achieve space-efficient and simple integration of the box with an input reservoir (e.g., a reservoir for storing fluid before the fluid passes through the box) and an output reservoir (e.g., a reservoir for storing fluid after the fluid passes through the box). Such systems rely on, for example, using a flexible tube to connect the box to an input reservoir and / or an output reservoir that can be provided, for example, in the form of a flexible bag. For example, these systems are not configured with respect to space efficiency, durability, simplicity, availability with existing laboratory equipment, and / or availability with a robotic control system.

[0006] Therefore, there is a need for a box with improved geometric configuration, improved space efficiency, improved durability, improved simplicity, improved usability with existing laboratory equipment, and / or improved usability with robotic control systems. The systems, methods, and techniques disclosed herein can meet one or more of the above needs.

[0007] Disclosed herein is a device for use with a system to deliver a payload into a cell. The device includes a box configured to receive and hold a contraction accommodating element, such as a microfluidic chip or a microfluidic filter. Unlike a box system that can be configured to be attached to an input reservoir by a flexible tube or by other fluidic components, the box disclosed herein may include an input reservoir physically integrated with the box itself. The input reservoir may be a reservoir configured to accommodate a cell suspension before the cell suspension flows through the contraction accommodating element. The box disclosed herein may be provided in the form of a top cover device, which is configured to be connected to and cover the opening of a container (such as a test tube, a culture tube, a sample tube, a centrifuge tube or the like). By using an integrated input reservoir, it is easy to integrate with existing laboratory devices (such as a pressure source that is easy to obtain). In addition, the timing and potential volume reduction of the embodiments described herein may be particularly useful in a research use environment, because the volume is small in a research use environment, and consumables can be used quickly via, for example, a large number of small volume experiments. The container (e.g., tube) to which the box can be attached as a top cover can serve as an output reservoir for containing a cell suspension after being processed by a constriction housing element. In some embodiments, when the box is in an attached position with the container (e.g., tube), a portion of the box can be disposed within the interior cavity of the container, for example, by protruding downward into the interior of a test tube. This configuration simplifies the retrieval of the tube container and can provide additional ease of use in a research environment.

[0008] Thus, when the box is attached to the open end of a container (such as a laboratory tube), the device formed by the combination of the box and the container can include an input reservoir, a receiving portion configured to receive and hold the contraction-containing element, and an output reservoir provided in the form of a container (such as a laboratory tube). All three components can be in fluid communication with each other to allow the cell suspension to flow from the input reservoir to the receiving portion (and into and out of the contraction-containing element contained therein), and then flow from the receiving portion to the container (e.g., a tube). After the cell suspension flows into the container (e.g., a tube), the box can be disengaged from the opening of the container, and the container (containing the processed cell suspension) can be used to further process the cell suspension, can be capped, and / or can be stored in any suitable manner. When the container is provided in the form of a standard laboratory tube, the tube can be easily used for downstream processing through one or more additional standard compatible devices, top covers, and / or storage devices.

[0009] Some embodiments are directed to a device for facilitating delivery of a payload to cells of a cell suspension, the device comprising: a container comprising an interior cavity and an opening; and a box configured to be connected to the opening of the container when in an assembled position relative to the container. The box comprises: an input reservoir configured to receive a cell suspension; and a receiving portion configured to receive a constriction housing element. The constriction housing element comprises a constriction configured to disrupt the membranes of cells of the cell suspension to facilitate delivery of a payload to the cells. The receiving portion is fluidically connected to the input reservoir to allow the cell suspension to flow from the input reservoir to the receiving portion, and the receiving portion is directly fluidically connected to a box outlet port to allow the cell suspension to flow from the receiving portion into the interior cavity of the container.

[0010] In some embodiments, when the cartridge is in the assembled position relative to the container, a portion of the receiving portion of the cartridge is disposed within the interior cavity of the container. In some embodiments, when the cartridge is in the assembled position relative to the container, a portion of the input reservoir of the cartridge is disposed within the interior cavity of the container.

[0011] In some embodiments, the device includes a covering configured to be releasably attached to the receiving portion of the box to hold the contraction housing element in place when the contraction housing element is received by the receiving portion. In some embodiments, the covering is configured to be slidably attached to and detached from the receiving portion of the box. In some embodiments, the covering is configured to slide in a direction perpendicular to the direction in which the layers of the contraction housing element are stacked on each other. In some embodiments, the covering is configured so that when the box is in the assembled position relative to the container, the inner wall of the container prevents the covering from detaching from the receiving portion.

[0012] In some embodiments, the box 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.

[0013] In some embodiments, the cassette includes a first O-ring disposed against a surface of the input reservoir cover, the first O-ring facing away from the interior of the input reservoir when the input reservoir cover is in the closed position. In some embodiments, the cassette includes a second O-ring disposed between the input reservoir cover and a main body portion of the cassette.

[0014] In some embodiments, the input reservoir cover comprises a filter configured to prevent backflow of the cell suspension. In some embodiments, the box has a total fluid throughput greater than 0.05 L / min. In some embodiments, the box is configured to allow the cell suspension to flow through the box at a pressure greater than 100 PSI.

[0015] Some embodiments are directed to a device for facilitating delivery of a payload to cells of a cell suspension. The device includes: a container including an interior cavity and an opening; and a box configured to be connected to the opening of the container when in an assembled position relative to the container, the box including: an input reservoir configured to receive a cell suspension; and a receiving portion. The device includes a constriction containing element. The constriction containing element includes a constriction configured to disrupt the membranes of cells of the cell suspension to facilitate delivery of a payload to the cells. The receiving portion of the box is fluidly connected to the input reservoir and the constriction containing element so as to allow the cell suspension to flow from the input reservoir through the receiving portion and into the constriction containing element, and so as to allow the cell suspension to flow out of the constriction containing element through the receiving portion and into the interior cavity of the container.

[0016] In some embodiments, the device comprises a gasket disposed between the receiving portion and the constriction containing element. In some embodiments, the constriction containing element comprises a microfluidic chip comprising a plurality of constrictions that disrupt the membrane of the cell. In some embodiments, the constriction containing element has a footprint of less than 5 mm x 5 mm.

[0017] In some embodiments, when the cartridge is in the assembled position relative to the container, a portion of the receiving portion of the cartridge is disposed within the interior cavity of the container. In some embodiments, when the cartridge is in the assembled position relative to the container, a portion of the input reservoir of the cartridge is disposed within the interior cavity of the container.

[0018] In some embodiments, the box further comprises a covering configured to be releasably attached to the receiving portion of the box to hold the contraction housing element in place when the contraction housing element is received by the receiving portion. In some embodiments, the covering is configured to be slidably attached to and detached from the receiving portion of the box. In some embodiments, the covering is configured to slide in a direction perpendicular to the direction in which the layers of the contraction housing element are stacked on each other. In some embodiments, the covering is configured so that when the box is in the assembled position relative to the container, the inner wall of the container prevents the covering from detaching from the receiving portion.

[0019] In some embodiments, the box 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.

[0020] In some embodiments, the cassette includes a first O-ring disposed against a surface of the input reservoir cover, the first O-ring facing away from the interior of the input reservoir when the input reservoir cover is in the closed position. In some embodiments, the cassette includes a second O-ring disposed between the input reservoir cover and a main body portion of the cassette.

[0021] In some embodiments, the input reservoir cover comprises a filter configured to prevent backflow of the cell suspension. In some embodiments, the box has a total fluid throughput greater than 0.05 L / min. In some embodiments, the box is configured to allow the cell suspension to flow through the box at a pressure greater than 100 PSI.

[0022] Some embodiments are directed to a cartridge for facilitating delivery of a payload to cells of a cell suspension, the cartridge comprising an input reservoir configured to receive the cell suspension, a connecting portion configured to connect to an opening to an interior cavity of a container, and a receiving portion configured to receive a constriction housing element. The constriction housing element comprises a constriction configured to disrupt the membranes of cells of the cell suspension to facilitate delivery of a payload to the cells. The receiving portion is fluidically connected to the input reservoir to allow the cell suspension to flow from the input reservoir to the receiving portion, and the receiving portion is fluidically connected to a cartridge outlet port to allow the cell suspension to flow out of the cartridge.

[0023] In some embodiments, when the cassette is connected in an assembled position relative to the container, a portion of the receiving portion of the cassette is disposed within the interior cavity of the container. In some embodiments, when the cassette is connected in an assembled position relative to the container, a portion of the input reservoir of the cassette is disposed within the interior cavity of the container.

[0024] In some embodiments, the box further comprises a covering configured to be releasably attached to the receiving portion of the box to hold the contraction accommodating element in place when the contraction accommodating element is received by the receiving portion. In some embodiments, the covering is configured to be slidably attached to and detached from the receiving portion of the box. In some embodiments, the covering is configured to slide in a direction perpendicular to the direction in which the layers of the contraction accommodating element are stacked on each other. In some embodiments, the covering is configured so that when the box is connected in an assembled position relative to the container, the inner wall of the container prevents the covering from detaching from the receiving portion.

[0025] In some embodiments, the cassette includes an input reservoir cover that is movable between an open position and a closed position to selectively expose and enclose an interior of the input reservoir. In some embodiments, the cassette includes a first O-ring disposed against a surface of the input reservoir cover, the first O-ring facing away from the interior of the input reservoir when the input reservoir cover is in the closed position. In some embodiments, the cassette includes a second O-ring disposed between the input reservoir cover and a main body portion of the cassette.

[0026] In some embodiments, the input reservoir cover includes a filter configured to prevent the cell suspension from flowing back. In some embodiments, the box has a total fluid throughput greater than 0.05 L / min. In some embodiments, the box is configured to allow the cell suspension to flow through the box at a pressure greater than 100 PSI. In some embodiments, the box has a total fluid throughput less than 0.5 L / min. In some embodiments, the box is configured to allow the cell suspension to flow through the box at a pressure less than 200 PSI.

[0027] In some embodiments, any one or more features, characteristics, or elements discussed above with respect to any embodiment may be incorporated into any other embodiment mentioned above or described elsewhere herein. In some embodiments, any one or more features, characteristics, or elements discussed elsewhere in this disclosure may be incorporated into any one or more embodiments mentioned above. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 A diagram illustrates a system for delivering a payload to a cell, according to some embodiments.

[0029] FIG. 2A to FIG. 2E The figures illustrate various views of an apparatus including a cartridge and a container for use in a system for delivering a payload to a cell, according to some embodiments.

[0030] FIG. 3A to FIG. 3B Figure shows a view of a cartridge for use in a system for delivering a payload to a cell, according to some embodiments.

[0031] FIG. 4A to FIG. 4B Figure illustrates a view of a chip and shim assembly for use with a cartridge used in a system for delivering a payload to a call, according to some embodiments. DETAILED DESCRIPTION

[0032] Described below are exemplary embodiments of cartridges for use in systems for partially or fully automated intracellular payload delivery, as well as associated devices, systems, methods, and techniques.

[0033] The following description sets forth exemplary systems, methods, techniques, parameters, and the like. However, it should be appreciated that such description is not intended to limit the scope of the present disclosure, but is provided as a description of exemplary embodiments.

[0034] definition

[0035] For the purpose of interpreting this specification, the following definitions shall apply and, where appropriate, terms used in the singular shall also include the plural and vice versa. If any definition set forth below conflicts with any document incorporated herein by reference, the definition set forth shall prevail.

[0036] As used herein, the term "pore" refers to an opening, including but not limited to a hole, a crack, a cavity, a pore, a fracture, a gap, or a perforation in a material. In some examples, (where specified and / or clear to one skilled in the art from this disclosure) the term refers to a pore in a surface of the present disclosure. In other examples, (where specified and / or clear to one skilled in the art from this disclosure) a pore may refer to a pore in a cell membrane.

[0037] As used herein, the term "filter" refers to a porous article that allows selective passage through the pores. In some examples, the term refers to a surface or membrane containing pores.

[0038] Although the description herein uses the terms first, second, etc. to describe various elements, these elements should not be limited by the terms. These terms are only used to distinguish one element from another.

[0039] For any of the structural and functional properties described herein, methods of determining such properties are known in the art.

[0040] All references cited herein, including patent applications and publications, are incorporated by reference in their entirety.

[0041] Cassettes for use in intracellular payload delivery systems

[0042] Figure 1 A schematic diagram of an intracellular payload delivery system 100 is shown.

[0043] In some embodiments, system 100 may share any one or more features with any of the systems described in WO 2019 / 126212 and / or WO 2020 / 210162, the entire contents of which are hereby incorporated by reference herein. In short, an intracellular payload delivery system can deliver a payload to a cell by forcing the cell to flow through a constriction (such as a narrow microfluidic channel or a narrow pore) thereby disrupting the cell's membrane and allowing the payload to enter the cell.

[0044] Although WO 2020 / 210162 describes a system for intracellular payload delivery, the system described in WO 2020 / 210162 relies on a box that is fluidly connected by a flexible tube and is configured to receive a flow of cell suspension from a preparation container; and the box is fluidly connected by a flexible tube and is configured to direct the flow of cell suspension to an output bag. As described herein, the box disclosed herein may differ from the box described in WO 2020 / 210162 in that the box described herein may be provided in the form of a top cover device configured to be connected to a container (such as a test tube), wherein the box may include an input reservoir formed as an integral part of the top cover device.

[0045] In some embodiments of system 100, a constriction (such as a narrow microfluidic channel or a narrow hole) can be provided in a microfluidic chip or filter (which can be referred to as a constriction-containing element) that can be fluidically attached to system 100. In some embodiments, a microfluidic chip or filter (or any other element including one or more constrictions configured to disrupt the membrane of a cell) can be provided and fluidically connected to a system such as system 100 via a cassette. A cassette can be any device that is configured to house an element including a constriction (such as a microfluidic chip or filter) and / or facilitate fluid connection of an element (e.g., a chip or filter) with another part of an intracellular payload delivery system (e.g., system 100).

[0046] like Figure 1 As shown, the system 100 can include a cartridge 300, a container 204, a cell suspension source 108, and a pressure source 110. The cartridge 300 and the container 204 can together form the device 200.

[0047] The box 300 can be formed as a top cover of the container 204 and can be configured to be attached to the container 204 by a threaded connection, a press connection, a snap-fit ​​connection, or any other suitable attachment method. When in an attached position relative to the container 204, the box 300 can cover the opening of the container 204. Figure 1 In the arrangement shown, the opening may be an opening in the top of the container 204 .

[0048] The container 204 can be any suitable container having one or more outer walls enclosing an interior cavity and having at least one opening leading to the interior cavity, wherein the opening is configured to cooperate with the box 300. In some embodiments, the container 204 can be a tube, such as a test tube, a culture tube, a sample tube, a centrifuge tube, or the like. In some embodiments, the container 204 can be an Eppendorf tube.

[0049] Box 300 may include an input reservoir and a receiving portion. The input reservoir may include a cavity formed in the box and configured to hold the cell suspension before the cell suspension flows through a constriction containing element (such as a microfluidic chip or a filter). The receiving portion may include a portion of box 300 that is configured to receive and hold the constriction containing element itself. The input reservoir and the receiving portion may be fluidly connected to each other, for example, by one or more internal channels, so that the cell suspension can flow from the input reservoir to the receiving portion and enter the constriction containing element contained in the receiving portion. After flowing through the constriction containing element, the cell suspension can flow out of the constriction containing element and return to the receiving portion. Then, the cell suspension can flow out of the box from the receiving portion and enter the cavity of the container, for example, by one or more internal channels.

[0050] The system 100 may also include a cell suspension source 108, which may include any source (e.g., a reservoir, a tank, a container, etc.) from which the cartridge 300 may receive a cell suspension. In some embodiments, an input opening leading to an input reservoir of the cartridge 300 may be configured to be fluidly connected to the cell suspension source 108 (e.g., by being selectively connectable and disconnectable) such that the cell suspension may flow from the cell suspension source 108 to the input reservoir of the cartridge 300.

[0051] The system 100 may also include a pressure source 110, which may include any source configured to apply pressure to the cell suspension within the input reservoir of the cartridge 300. In some embodiments, the pressure source 110 may be provided as part of the same fluid source mechanism as the cell suspension source 108. In some embodiments, the pressure source 110 may be configured to connect (e.g., by being selectively connectable and disconnectable) the input reservoir of the cartridge 300 so as to apply pressurized gas, pressurized liquid, or / and mechanical pressure to the cell suspension within the input reservoir of the cartridge 300. The pressure applied to the cell suspension within the input reservoir of the 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 receiving portion of the cartridge 300. In some embodiments, the pressure source 110 may share one or more characteristics of the pressure control module and / or other pressure control components described in WO 2019 / 126212. As noted above, WO 2019 / 126212 is incorporated herein by reference in its entirety for all purposes.

[0052] An exemplary embodiment of an improved cassette for use in a system for intracellular payload delivery, such as cassette 300 for use in system 100 , is described below.

[0053] FIG. 2A to FIG. 2E Various views of an apparatus 200 including a cartridge 300 and a container 204 for use in a system 100 for delivering a payload to a cell are shown, according to some embodiments.

[0054] The box 300 can be configured to accommodate one or more constriction-containing elements, such as a constriction filter (including one or more constriction holes) or a constriction microfluidic chip (including one or more constriction microfluidic channels). (Constriction filters according to some embodiments are disclosed in application No. WO / 2017 / 041050A1, which is hereby incorporated by reference in its entirety.) It should be noted that in some embodiments, the constriction microfluidic channel or constriction hole can be simply referred to as a "constriction" or "cell deformation constriction". The constriction-containing element can be any component, device, part or the like having a channel, passage or other opening (e.g., a constriction) whose diameter is smaller than the cells of the cell suspension passing through the element, so that forcing the cells through the opening under pressure causes a disturbance in the membrane of the cells because the cells are constricted by the opening. In some embodiments, the device 200 and / or the box 300 can include an integrated constriction filter or microfluidic channel configured to constrict the cells, while in some embodiments, the box 200 can be configured to accommodate different elements (e.g., chips or filters) that themselves include constriction holes or constriction microfluidic channels. In either case, device 200 and cartridge 300 can define a portion of a flow path of a system (such as system 100) for delivering a payload to cells such that a cell suspension can flow toward and into cartridge 300, and such that the cell suspension can then flow through and out of cartridge 300 and toward and into container 204 (and / or into any other suitable downstream flow path component).

[0055] Figure 2A A perspective view of the device 200 is shown. Figure 2B An exploded perspective view of the device 200 is shown. Figure 2C A first side view of device 200 is shown. Figure 2D A second side view of the device 200 is shown. Figure 2C The first side view is shown rotated 90 degrees. Figure 2E A top view of the device 200 is shown. FIG. 2A to FIG. 2E In each of the views of FIG. 3 , the device 200 is shown with the chip and gasket assembly inserted into the receiving portion 306 of the cassette 300 .

[0056] exist FIG. 2A to FIG. 2C In each view in FIG. 3 , portions of the box 300 (e.g., the box body 302) and the container 202 are shown as partially transparent. In some embodiments, some or all portions of the box 300 and / or the container 202 may be transparent, translucent, or opaque.

[0057] exist FIG. 2A to FIG. 2EIn the illustrated embodiment, the container 204 is in the form of a tube, such as a test tube, a culture tube, a sample tube, a centrifuge tube or the like. In some embodiments, the container 204 can be an Eppendorf tube. Like this, the container 204 can be an off-the-shelf component with additional configuration to accommodate the chip.

[0058] In some embodiments, container 204 may include one or more components made of metal, plastic, polymer and / or glass. In some embodiments, container 204 may include one or more components made of polycarbonate, polypropylene and / or polymethyl methacrylate.

[0059] In some embodiments, the lumen of the container 204 can have a volume greater than or equal to 1 mL, 1.5 mL, 2 mL, 2.5 mL, 3 mL, or 3.5 mL. In some embodiments, the lumen of the container 204 can have a volume less than or equal to 1 mL, 1.5 mL, 2 mL, 2.5 mL, 3 mL, or 3.5 mL.

[0060] like FIG. 2A to FIG. 2E As further shown, the box 300 (reference FIG. 3A to FIG. 3B The cartridge 300 (shown and described in further detail) can be configured to mate with the opening 205 of the container 204, for example by taking the form of a top cover of the container 204. In some embodiments, the cartridge 300 can be configured to mate with the opening 205 of the container 204 by a press connection, a snap connection, a twist lock connection, or any other suitable connection. In some embodiments, the cartridge 300 can be configured to be held against the opening 205 of the container 204 by one or more tube handling devices (such as a robotic arm) during processing of the cell suspension by the cartridge 300.

[0061] As described above, unlike the box system that can be configured to be attached to the input reservoir by a flexible tube or by other fluid control components, the box disclosed herein may include an input reservoir physically integrated with the box itself. The input reservoir may be a reservoir configured to hold a cell suspension before the cell suspension flows through the contraction accommodating element. The box disclosed herein may be provided in the form of a top cover device, which is configured to be connected to and cover the opening of a container (such as a test tube, a culture tube, a sample tube, a centrifuge tube or the like). By using an integrated input reservoir, it is easy to integrate with existing laboratory devices (such as a pressure source that is easy to obtain). In addition, the timing and potential volume reduction of the embodiments described herein may be particularly useful in a research use environment, because the volume is small in a research use environment, and consumables can be used quickly via, for example, a large number of small volume experiments. The container (e.g., tube) to which the box can be attached as a top cover can serve as an output reservoir, which is used to hold a cell suspension after being processed by the contraction accommodating element. In some embodiments, when the box is in an attached position with the container (e.g., tube), a portion of the box can be disposed within the interior cavity of the container, such as by protruding downward into the interior of the test tube. This configuration simplifies the retrieval of the tube container and can provide additional ease of use in a research environment.

[0062] Thus, when the box is attached to the open end of a container (such as a laboratory tube), the device formed by the combination of the box and the container can include an input reservoir, a receiving portion configured to receive and hold the contraction-containing element, and an output reservoir provided in the form of a container (such as a laboratory tube). All three components can be in fluid communication with each other to allow the cell suspension to flow from the input reservoir to the receiving portion (and into and out of the contraction-containing element contained therein), and then flow from the receiving portion to the container (e.g., a tube). After the cell suspension flows into the container (e.g., a tube), the box can be disengaged from the opening of the container, and the container (containing the processed cell suspension) can be used to further process the cell suspension, can be capped, and / or can be stored in any suitable manner. When the container is provided in the form of a standard laboratory tube, the tube can be easily used for downstream processing through one or more additional standard compatible devices, top covers, and / or storage devices.

[0063] like Figure 2A and FIG. 2C to FIG. 2EAs shown, the box 300 can be configured so that when it is assembled with the container 204 by being placed in the assembly position, the box body 302 extends downward into the interior cavity of the container 204. In some embodiments, the width of a portion of the box body 302 can be less than the inner diameter of the interior cavity of the container 204, so that the portion of the box body 302 can extend downward into the interior cavity of the container 204. In some embodiments, the box body 302 can include a rim portion that is wider than the portion that extends into the interior cavity of the container 204, so that the rim portion can be located outside (e.g., at the top of) the opening 205 of the container 204, while a portion of the box body 302 extends into the interior cavity of the container 204. In some embodiments, when in the assembly position with respect to the container 204, the box 300 can extend into the interior cavity of the container 204 by an amount greater than or equal to 30%, 40%, 50%, 60%, or 70% of the total height dimension of the interior cavity of the container 204 (along the direction in which the box 300 extends into the interior cavity). In some embodiments, the cartridge 300 may extend into the interior cavity of the container 204 when in the assembled position with respect to the container 204 by an amount less than or equal to 30%, 40%, 50%, 60%, or 70% of the total height dimension of the interior cavity of the container 204 (in the direction in which the cartridge 300 extends into the interior cavity). In some embodiments, the cartridge 300 may extend into the interior cavity of the container 204 by greater than or equal to 0.5 cm, 1 cm, 1.5 cm, 2 cm, or 2.5 cm when in the assembled position with respect to the container 204. In some embodiments, the cartridge 300 may extend into the interior cavity of the container 204 by less than or equal to 0.5 cm, 1 cm, 1.5 cm, 2 cm, or 2.5 cm when in the assembled position with respect to the container 204.

[0064] In some embodiments, when in an assembled position with respect to the container 204, greater than or equal to 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the cartridge 300 in terms of height may extend into the interior cavity of the 204. In some embodiments, when in an assembled position with respect to the container 204, less than or equal to 50%, 60%, 70%, 80%, 90%, 95%, or 99% of the cartridge 300 in terms of height may extend into the interior cavity of the 204.

[0065] Specific as Figure 2B As shown, in some embodiments, the cartridge 300 can include a cartridge body 302 including an input reservoir 304 and a receiving portion 306. The input reservoir 304 can be configured to receive and hold a cell suspension before the cell suspension flows from the input reservoir 304 to the receiving portion 306.

[0066] In some embodiments, the input reservoir can have a tapered shape that narrows at the bottom so that fluid can flow out of the bottom of the input reservoir 304 into one or more channels under the action of gravity and / or pressure (e.g., liquid pressure, gas pressure, and / or mechanical pressure). In some embodiments, the input reservoir 306 can have an internal volume less than or equal to 1 mL, 1.5 mL, 2 mL, 2.5 mL, or 3 mL. In some embodiments, the input reservoir 306 can have an internal volume greater than or equal to 1 mL, 1.5 mL, 2 mL, 2.5 mL, or 3 mL.

[0067] The receiving portion 306 may be configured to receive and hold a constriction housing component, such as a chip and gasket assembly 400 (see FIG. 4A to FIG. 4B 204 ), and allows the cell suspension to flow from the input reservoir 304 into the receiving portion and into the constriction containment element through one or more channels in the receiving portion. After the cell suspension has flowed through the constriction containment element, the suspension may flow back into the one or more channels of the receiving portion. The one or more channels of the receiving portion may lead to the exterior of the cartridge 300 so that the cell suspension may flow out of the cartridge 300 and into the interior cavity of the container 204.

[0068] The cassette 300 may include a receiving portion cover 308 that may be configured to cover the receiving portion 306 and hold the constriction housing element in place in the receiving portion 306 .

[0069] The receiving portion covering 308 can be an element configured to be placed next to one or more constriction containing elements to press the one or more constriction containing elements toward the receiving portion 306 and / or otherwise hold the one or more constriction containing elements in place. In some embodiments, the receiving portion covering 308 can be configured to apply an inward force to the one or more constriction containing elements to press them toward the receiving portion 306 through one or more springs or other compressible or deformable components (such as one or more rubber O-rings or gaskets). In some embodiments, the receiving portion covering 308 can be configured to press against the surface of one or more constriction containing elements. In some embodiments, the receiving portion covering 308 can be used to ensure that one or more constriction containing elements do not delaminate under the pressure of a fluid forced through them; by pressing the receiving portion covering 308 against one face of the constriction containing element under the action of force, the constriction containing element can be prevented from delaminating.

[0070] The receiving portion covering 308 can be attachable and removable to the receiving portion 306, for example, by a sliding connection, a threaded connection, a hinge connection, a protrusion and seam connection, a locking mechanism, by one or more screws, by one or more cams, or in any other suitable manner, so that the receiving portion covering 308 can be removed, for example, to replace the gasket 404 and / or the retraction portion accommodating element 402.

[0071] In some embodiments, a sliding connection (such as FIG. 2A to FIG. 2E The connection shown) can allow the receiving portion cover 308 to slide laterally along the receiving portion 306 (in Figure 2B 306) such that the lip of the receiving portion cover 308 can slide over a corresponding lip, protrusion, groove or tooth of the receiving portion 306. In some embodiments, a removable cover configured to slide onto and off the body of the receiving portion can be configured to completely surround the receiving portion, thereby avoiding the need for interlocking grooves or teeth or the like in some embodiments.

[0072] A sliding connection such as this can be removed with minimal lateral force (e.g., force in the sliding direction), but can provide great strength in a direction perpendicular to the sliding direction and extending away from the receiving portion where the covering is placed. Therefore, although a covering with a sliding connection can be easily removed by hand, it can still provide better pressure durability than other connection mechanisms that can be used to manually assemble the box (such as attaching the covering with threaded components). In some embodiments, in addition to or in lieu of one or more removable coverings, the box can be configured to securely accommodate one or more constriction-receiving elements without the use of removable coverings.

[0073] In some embodiments, the receiving portion cover 308 can be configured to be slidably removed from the receiving portion 306 such that the receiving portion cover cannot be slidably removed from the receiving portion 306 when the cartridge 300 is in an assembled position with respect to the container 204. FIG. 2A to FIG. 2E As shown, when the box 300 is assembled with the container 204 so that the box body 302 extends downwardly into the interior cavity of the container 204, the walls of the container 204 can prevent the receiving portion cover 308 from sliding off either side of the receiving portion 306. This arrangement can help ensure that the receiving portion cover 308 is not accidentally removed from the receiving portion during operation.

[0074] The cartridge 300 can be configured so that a cell suspension fluid can be introduced through an opening (such as Figure 2BThe top opening 305 shown enters the input reservoir 304. In some embodiments, the input reservoir 304 can have an open top so that the cell suspension can flow into the input reservoir 304 from the top.

[0075] In some embodiments, the box 300 can include a cover 326 that can cover the opening of the input reservoir 304. The cover 326 can be configured to cooperate with the opening of the input reservoir 304 by a press connection, a snap connection, a twist lock connection, or any other suitable connection. The cover 326 can be a foldable cover including an upper portion 327 and a lower portion 329, wherein the upper portion 327 can be folded upward to an open position to allow access to the input reservoir 304, and then folded downward to a closed position to close the cell suspension in the input reservoir 304. In some embodiments, the cover 326 can be placed in a closed position before applying pressure to the cell suspension in the input reservoir 304. In some embodiments, the cover 326 can include a filter 324, which can be configured to prevent liquid (e.g., cell suspension) from flowing back from the box to the cell suspension source and / or the pressure source. Air pressure can be supplied, for example, from the pressure source above the filter 324.

[0076] like Figure 2B and Figure 2C As shown specifically, the cartridge 300 may include two or more O-rings, including an O-ring 320 and an O-ring 322. The O-ring 320 may be disposed between the cover 326 and the cartridge body 302 to form a seal therebetween. The O-ring 322 may be disposed on a side of the cover 326 opposite the cartridge 302 and may form a seal between the cover 326 and another component of the system 100, such as the cell suspension source 108 and / or the pressure source 110. In some embodiments, the O-ring 322 may be pressed against another component of the system 100 (e.g., by a robotic arm) to form a seal and allow positive pressure to be applied to the cell suspension within the input reservoir 304.

[0077] FIG. 3A to FIG. 3B The figure shows a view of a cartridge 300 for use in a system 100 for delivering a payload to a cell, according to some embodiments.

[0078] Figure 3A A side view of the cartridge 300 is shown. Figure 3B A close-up side view of the lower portion of the device 300 is shown, with focus on the receiving portion 306 and the receiving portion cover 308. Figure 3A and Figure 3B In both, the cassette is shown with the chip and gasket assembly inserted into cassette 300.

[0079] exist Figure 3A and Figure 3B, portions of the box 300 (e.g., the box body 302) are shown as partially transparent. In some embodiments, some or all portions of the box 300 may be transparent, translucent, or opaque.

[0080] Figure 3A and Figure 3B A view of an internal channel formed within the cartridge body 302 that facilitates the flow of a cell suspension through the cartridge 300 is provided.

[0081] As shown, the input channel 310 fluidly connects the input reservoir 304 to the receiving portion 306. In the illustrated arrangement, the input channel 310 has an initial vertical portion and a subsequent second horizontal portion, starting at the outlet of the input reservoir 304 and terminating at a portion of the receiving portion 306 that is configured to hold a gasket for a constriction housing element (with respect to FIG. 4A to FIG. 4B 1.5 mm, 1.5 mm, 2 mm, or 2.5 mm. In some embodiments, all or a portion of the input channel 310 can have a diameter less than or equal to 0.05 mm, 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm, or 2.5 mm. In some embodiments, all or a portion of the input channel 310 can have a diameter greater than or equal to 0.05 mm, 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm, or 2.5 mm.

[0082] As shown, the output channel 312 begins at a portion of the receiving portion 306 that is configured to hold a gasket for the constriction containment element and terminates at an opening to the exterior of the cartridge 300. Thus, a cell suspension that has passed through the constriction containment element can flow through the output channel 312 and out of the cartridge 300, for example dripping downward under the action of gravity to be collected in a container (e.g., container 204) into which the cartridge 300 is inserted. In some embodiments, all or part of the output channel 312 can have a diameter less than or equal to 0.05 mm, 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm, or 2.5 mm. In some embodiments, all or part of the output channel 312 can have a diameter greater than or equal to 0.05 mm, 0.1 mm, 0.5 mm, 1 mm, 1.5 mm, 2 mm, or 2.5 mm.

[0083] In some embodiments, the height of the box 300 (e.g., in Figure 2A In the vertical direction shown in the figure, the height of the box 300 can be less than or equal to 0.5mm, 0.6mm, 0.75mm, 1mm, 5mm, 1cm, 5cm or 10cm. In some embodiments, the height of the box 300 can be greater than or equal to 0.5mm, 0.6mm, 0.75mm, 1mm, 5mm, 1cm, 5cm or 10cm.

[0084] In some embodiments, the width of the box 300 (e.g., in Figure 2A The width of the box 300 (in the direction extending horizontally in the middle) can be less than or equal to as measured at the portion of the box 300 extending into the interior cavity of the container 204. In some embodiments, when in an assembled position relative to the container 204, the width of the box 300 can be less than or equal to as measured at the portion of the box 300 extending into the interior cavity of the container 204.

[0085] FIG. 4A to FIG. 4B The figure shows a view of a chip and spacer assembly 400 for use with a cartridge used in a system 100 for delivering a payload to a cell, according to some embodiments. The chip and spacer assembly 400 can be used with FIG. 2A to FIG. 2D , Figure 3A and Figure 3B The chip and gasket assemblies shown are identical. FIG. 4A to FIG. 4B An exemplary microfluidic chip including a plurality of constrictions configured to induce perturbations in cell membranes is shown, but it will be appreciated in light of this disclosure that a microfluidic filter configured to induce perturbations in cell membranes may additionally or alternatively be used.

[0086] Figure 4A A perspective view of a chip and gasket assembly 400 is shown. Figure 4B A plan view of the chip and gasket assembly 400 is shown with the gasket 404 in the foreground.

[0087] exist Figure 3A and Figure 3B , portions of the box 300 (e.g., the box body 302) are shown as partially transparent. In some embodiments, some or all portions of the box 300 may be transparent, translucent, or opaque.

[0088] like Figure 4A Specifically shown, the chip and spacer assembly 400 may include a chip 402 and a spacer 404, which may be positioned adjacent to each other in a layered arrangement. The chip 402 itself may include a shrink layer 402a and a cover layer 402b.

[0089] The contraction layer 402a may be a layer where one or more cell-deforming contractions are formed (eg, by etching or machining into the layer). In some embodiments, the contraction layer 402a may be formed of silicon.

[0090] The cover layer 402b can be placed against the contraction layer 402a so as to cover one or more cavities and / or contractions formed in the contraction layer 402a. Thus, the cover layer 402b can form a "lid" on top of the cover layer 402a and seal the volume area through which the cell suspension flows in the contraction layer 402a. Figure 4A and Figure 4B As shown, the cover layer 402a may include a chip inlet channel 408 and a chip outlet channel 410, each of which may be formed as a through hole in the cover layer 402a to allow a cell suspension to flow through the channel. The cell suspension may flow into the chip 402 and enter the shrink layer 402a through the chip inlet channel 408, and may flow out of the shrink layer 402a and enter the chip outlet channel 410 and flow out of the chip 402. In some embodiments, the cover layer 402b may be formed of glass. The cover layer 402b may be bonded to the shrink layer 402a by any suitable means (such as by an adhesive).

[0091] In some embodiments, one or both of the chip inlet channel 408 and the chip outlet channel 410 can have a diameter less than or equal to 0.2 mm, 0.4 mm, 0.6 mm, 0.8 mm, 1 mm, or 1.2 mm. In some embodiments, one or both of the chip inlet channel 408 and the chip outlet channel 410 can have a diameter greater than or equal to 0.2 mm, 0.4 mm, 0.6 mm, 0.8 mm, 1 mm, or 1.2 mm.

[0092] Gasket 404 may be configured to seat against cover layer 402a. Gasket 404 may provide a channel fluidly connecting chip 402 to a channel of receiving portion 306, thereby forming a seal with receiving portion 306 and allowing fluid to flow from receiving portion 306 to chip 402 and back to receiving portion 306.

[0093] like Figure 4A and Figure 4B As shown, the gasket 404 may include a gasket inlet channel 406 and a gasket outlet channel 412, each of which may be formed as a through hole in the gasket 404 to allow the cell suspension to flow through the channel. The cell suspension may flow into the chip 402 through the gasket inlet channel 406, and may flow out of the chip 402 into and through the gasket outlet channel 412.

[0094] In some embodiments, one or both of the shim inlet channel 406 and the shim outlet channel 412 can have a cross-sectional dimension (eg, fluid diameter) of 0.1 mm to 2.0 mm.

[0095] In some embodiments, gasket 404 may be formed from one or more silicone, rubber, and / or thermoplastic elastomers.

[0096] Thus, the cell suspension may flow from the input reservoir 304 to the input channel 310, from the input channel 310 to the gasket inlet channel 406, from the gasket inlet channel 406 to the chip inlet channel 408, from the chip inlet channel 408 to the contraction layer 402a, from the contraction layer 402a to the chip outlet channel 410, from the chip outlet channel 410 to the gasket outlet channel 412, from the gasket outlet channel 412 to the output channel 312, and from the output channel 312 out of the cartridge 300 and into the interior cavity of the container 204. Buffer or other fluids may also flow along the same or similar flow paths.

[0097] In some embodiments, in addition to or as an alternative to using a gasket such as gasket 414, other sealing options may be used to create a seal for the fluid connection between the box and the constriction containing element; for example, O-rings, overmolding, chemical bonding, and / or mechanical interlocking may be used.

[0098] In some embodiments, chip 402 may have a length and / or width less than or equal to 50 mm, 25 mm, 10 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm. In some embodiments, chip 402 may have a length and / or width greater than or equal to 50 mm, 25 mm, 10 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm. In some embodiments, chip 402 may have a footprint of 4.1 mm×4.1 mm.

[0099] In some embodiments, chip 402 may have a thickness less than or equal to 0.1 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, or 5 mm. In some embodiments, chip 402 may have a thickness less than or equal to 0.1 mm, 0.5 mm, 1 mm, 2 mm, 3 mm, or 5 mm.

[0100] In some embodiments, the shrink layer 402a can have a thickness less than or equal to 0.2 mm, 0.4 mm, 0.6 mm, 0.8 mm, 1 mm, 1.5 mm, or 2 mm. In some embodiments, the shrink layer 402a can have a thickness less than or equal to 0.2 mm, 0.4 mm, 0.6 mm, 0.8 mm, 1 mm, 1.5 mm, or 2 mm.

[0101] In some embodiments, the cover layer 402b can have a thickness less than or equal to 0.2 mm, 0.4 mm, 0.6 mm, 0.8 mm, 1 mm, 1.5 mm, or 2 mm. In some embodiments, the cover layer 402b can have a thickness less than or equal to 0.2 mm, 0.4 mm, 0.6 mm, 0.8 mm, 1 mm, 1.5 mm, or 2 mm.

[0102] In some embodiments, the width of the constriction (e.g., constriction channel or constriction hole) of the constriction accommodating element (e.g., chip 402) can be less than or equal to 0.25 μm, 0.5 μm, 1 μm, 5 μm, 10 μm, 20 μm, or 50 μm. In some embodiments, the width of the constriction (e.g., constriction channel or constriction hole) of the constriction accommodating element (e.g., chip 402) can be greater than or equal to 0.25 μm, 0.5 μm, 1 μm, 5 μm, 10 μm, 20 μm, or 50 μm.

[0103] In some embodiments, the length of the constriction (e.g., constriction channel or constriction hole) of the constriction accommodation element (e.g., chip 402) can be less than or equal to 1 μm, 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, or 100 μm. In some embodiments, the length of the constriction (e.g., constriction channel or constriction hole) of the constriction accommodation element (e.g., chip 402) can be greater than or equal to 1 μm, 5 μm, 10 μm, 20 μm, 30 μm, 40 μm, 50 μm, or 100 μm.

[0104] In some embodiments, the depth of the constriction (e.g., constriction channel or constriction hole) of the constriction accommodation element (e.g., chip 402) can be less than or equal to 10 μm, 15 μm, 20 μm, 50 μm, 80 μm, 100 μm, or 200 μm. In some embodiments, the depth of the constriction (e.g., constriction channel or constriction hole) of the constriction accommodation element (e.g., chip 402) can be greater than or equal to 10 μm, 15 μm, 20 μm, 50 μm, 80 μm, 100 μm, or 200 μm.

[0105] In some embodiments, the spacer 404 may have a length and / or width less than or equal to 50 mm, 25 mm, 10 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm. In some embodiments, the spacer 404 may have a length and / or width greater than or equal to 50 mm, 25 mm, 10 mm, 5 mm, 4 mm, 3 mm, 2 mm, or 1 mm. In some embodiments, the spacer 404 may have a footprint of 4.1 mm×4.1 mm.

[0106] In some embodiments, the spacer 404 can have a thickness less than or equal to 0.25 mm, 0.5 mm, 0.75 mm, 1 mm, 1.25 mm, 1.5 mm, 1.75 mm, 2 mm, or 2.25 mm. In some embodiments, the spacer 404 can have a thickness less than or equal to 0.25 mm, 0.5 mm, 0.75 mm, 1 mm, 1.25 mm, 1.5 mm, 1.75 mm, 2 mm, or 2.25 mm.

[0107] In some embodiments, multiple constriction containment elements in 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 containment elements in the cartridge 300 may be arranged in parallel, such that as the fluid flows through the cartridge 300, the flow path in the cartridge 300 bifurcates into multiple parallel segments, which may then reconverge before flowing out of the cartridge 300. In some embodiments, three or more constriction containment elements may be arranged in the cartridge 300, such that one or more of the elements are connected in series with another constriction containment element, and one or more of the elements are connected in parallel with another constriction containment element.

[0108] In some embodiments, the cartridge 300 can be configured to receive blank placeholder elements in place of functional constriction containment elements, wherein the blank placeholder elements may not contain any channels or holes, or may be otherwise configured to not allow flow through the portion of the cartridge 300 that receives the placeholder elements. By using blank placeholder elements, the cartridge 300 can allow fluid to flow through a smaller number of constriction containment elements at a time, or through only one constriction containment element at a time, so that the system does not need to be used at full capacity of the constriction containment elements at all times.

[0109] In some embodiments, the device 200 can be configured to force fluid through the cartridge 300 at a pressure less than or equal to 1, 5, 10, 25, 50, 75, 100, 125, 150, or 200 PSI. In some embodiments, the device 200 can be configured to force fluid through the cartridge 300 at a pressure greater than or equal to 1, 5, 10, 25, 50, 75, 100, 125, 150, or 200 PSI. In some embodiments, the device 200 can be configured for use with constriction containment elements (e.g., constriction containment elements), each of which can individually (e.g., on a "per chip" basis) provide a throughput of less than or equal to 50, 100, 150, 200, 250, 300, or 400 mL of red blood cell suspension per minute. In some embodiments, device 200 can be configured for use with constriction containment elements (e.g., constriction containment elements), each of which can individually (e.g., on a "per chip" basis) provide a throughput of greater than or equal to 50, 100, 150, 200, 250, 300, or 400 mL of red blood cell suspension per minute.

[0110] In some embodiments, the device 200 can be configured for use with a contraction containing element (e.g., a contraction containing element), each of which can individually (e.g., on a "per chip" basis) provide a throughput of less than or equal to 25, 50, 75, 100, 125, 150, or 200 mL of a peripheral blood mononuclear cell suspension per minute. In some embodiments, the device 200 can be configured for use with a contraction containing element (e.g., a contraction containing element), each of which can individually (e.g., on a "per chip" basis) provide a throughput of greater than or equal to 25, 50, 75, 100, 125, 150, or 200 mL of a peripheral blood mononuclear cell suspension per minute.

[0111] In some embodiments, the device 200 can have a total fluid throughput (e.g., including all constriction containment elements contained in the cartridge 300) of less than or equal to 0.01 L / min, 0.05 L / min, 0.1 L / min, 0.25 L / min, 0.5 L / min, or 1 L / min. In some embodiments, the device 200 can have a total fluid throughput (e.g., including all constriction containment elements contained in the cartridge 300) of greater than or equal to 0.01 L / min, 0.05 L / min, 0.1 L / min, 0.25 L / min, 0.5 L / min, or 1 L / min.

Claims

1. A device for facilitating delivery of a payload to cells in a cell suspension, the device comprising: a container comprising an interior cavity and an opening; as well as A box configured to be connected to the opening of the container when in an assembled position relative to the container, the box comprising: an input reservoir configured to receive a cell suspension; as well as A receiving portion configured to receive a constriction housing element, wherein the constriction housing element includes a constriction configured to disrupt membranes of cells of the cell suspension to facilitate delivery of a payload to the cells, wherein the receiving portion is fluidically connected to the input reservoir to allow the cell suspension to flow from the input reservoir to the receiving portion, and wherein the receiving portion is directly fluidically connected to a cartridge outlet port to allow the cell suspension to flow from the receiving portion into the interior cavity of the container.

2. The apparatus of claim 1, wherein a portion of the receiving portion of the cartridge is disposed within the interior cavity of the container when the cartridge is in the assembled position relative to the container.

3. The apparatus of claim 1, wherein a portion of the input reservoir of the cartridge is disposed within 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 covering configured to be releasably attached to the receiving portion of the box to hold the retraction housing element in place when the retraction housing element is received by the receiving portion.

5. The apparatus of claim 4, wherein the cover is configured to be slidably attached to and detached from the receiving portion of the box.

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

7. The apparatus of claim 4, wherein the cover is configured such that when the box is in the assembled position relative to the container, an inner wall of the container prevents the cover from being disengaged from the receiving portion.

8. The device of claim 1, wherein the box 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.

9. The apparatus of claim 8, wherein the cartridge comprises a first O-ring seated against a surface of the input reservoir cover, the first O-ring facing away from the interior of the input reservoir when the input reservoir cover is in the closed position.

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

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

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

13. The apparatus 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. A device for facilitating delivery of a payload to cells in a cell suspension, the device comprising: a container comprising an interior cavity and an opening; A box configured to be connected to the opening of the container when in an assembled position relative to the container, the box comprising: an input reservoir configured to receive a cell suspension; and receiving part; and a constriction containing element, wherein the constriction containing element comprises a constriction configured to disrupt membranes of cells of the cell suspension to facilitate delivery of a payload to the cells; The receiving portion of the box is fluidly connected to the input fluid reservoir and the constriction containing element so as to allow the cell suspension to flow from the input fluid reservoir through the receiving portion and into the constriction containing element, and so as to allow the cell suspension to flow out of the constriction containing element through the receiving portion and into the internal cavity of the container.

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

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

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

18. The apparatus of claim 14, wherein a portion of the receiving portion of the cartridge is disposed within the interior cavity of the container when the cartridge is in the assembled position relative to the container.

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

20. The apparatus of claim 14, wherein the box further comprises a covering configured to be releasably attached to the receiving portion of the box to hold the retraction receiving element in place when the retraction receiving element is received by the receiving portion.

21. The apparatus of claim 20, wherein the cover is configured to be slidably attached to and detached from the receiving portion of the box.

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

23. The apparatus of claim 20, wherein the cover is configured such that when the box is in the assembled position relative to the container, an inner wall of the container prevents the cover from being disengaged from the receiving portion.

24. The device of claim 14, wherein the box includes an input reservoir cover, which is movable between an open position and a closed position to selectively expose and enclose the interior of the input reservoir.

25. The apparatus of claim 24, wherein the cartridge includes a first O-ring seated against a surface of the input reservoir cover, the first O-ring facing away from the interior of the input reservoir when the input reservoir cover is in the closed position.

26. The apparatus of claim 24, wherein the cartridge comprises a second O-ring disposed between the input reservoir cover and a main body portion of the cartridge.

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

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

29. The apparatus 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. 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 connecting portion configured to connect to an opening to the interior cavity of the container; as well as a receiving portion configured to receive a constriction housing element, wherein the constriction housing element includes a constriction configured to disrupt membranes of cells of the cell suspension to facilitate delivery of a payload to the cells, wherein the receiving portion is fluidly connected to the input reservoir to allow the cell suspension to flow from the input reservoir to the receiving portion, and wherein the receiving portion is fluidly connected to a cartridge outlet port to allow the cell suspension to flow out of the cartridge.

31. The cartridge of claim 30, wherein a portion of the receiving portion of the cartridge is disposed within the interior cavity of the container when the cartridge is connected in an assembled position relative to the container.

32. The cartridge of claim 30, wherein a portion of the input reservoir of the cartridge is disposed within the interior cavity of the container when the cartridge is connected in an assembled position relative to the container.

33. The box of claim 30, wherein the box further comprises a covering configured to be releasably attached to the receiving portion of the box to hold the retraction receiving element in place when the retraction receiving element is received by the receiving portion.

34. The case of claim 33, wherein the cover is configured to be slidably attached to and detached from the receiving portion of the case.

35. The box 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 each other.

36. The box of claim 33, wherein the cover is configured so that when the box is connected in an assembled position relative to the container, an inner wall of the container prevents the cover from being separated from the receiving portion.

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

38. The cartridge of claim 37, wherein the cartridge includes a first O-ring seated against a surface of the input reservoir cover, the first O-ring facing away from the interior of the input reservoir when the input reservoir cover is in the closed position.

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

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

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

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

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

46. ​​The apparatus 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. The apparatus 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. 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.

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