Electroporation device and method

Abstract

Electroporation is a technique that applies an electric field to cells to increase the permeability of the cell membrane. This allows for the introduction of chemicals, drugs, and / or macromolecules, such as proteins and nucleic acids, into the cells. Embodiments relate to an electroporation device. The electroporation device includes a plurality of chambers configured to store a plurality of cells during an electroporation process, a plurality of electrodes configured to generate a plurality of electric fields within the plurality of chambers during the electroporation process, each of the plurality of electric fields corresponding to one of the plurality of chambers, a flow channel configured to transport the plurality of cells during a cell collection process after the electroporation process, and a plurality of valves connecting the plurality of chambers to the flow channel.

Description

[0001] Cross-references to related applications

[0002] This application claims priority to U.S. Provisional Application No. 62 / 933,717, filed November 11, 2019, pursuant to 35 USC §119(e). This application also claims priority to U.S. Provisional Application No. 62 / 940,032, filed November 25, 2019, pursuant to 35 USC §119(e). Both U.S. Provisional Patent Application Nos. 62 / 933,717 and 62 / 940,032 are incorporated herein by reference in their entirety. Background Technology

[0003] Electroporation is a technique that applies an electric field to cells, thereby increasing the permeability of the cell membrane. This allows the introduction of drugs, chemicals, and / or macromolecules (such as proteins and nucleic acids (such as various forms of DNA and RNA)) into cells. Electroporation can also be called electrotransfer. Summary of the Invention

[0004] Generally, in one aspect, embodiments relate to an electroporation apparatus. The electroporation apparatus includes: a plurality of chambers configured to store a plurality of cells during an electroporation process; a plurality of electrodes configured to generate a plurality of electric fields within the plurality of chambers during the electroporation process, each of the plurality of electric fields corresponding to one of the plurality of chambers; a flow channel configured to transport the plurality of cells during a cell collection process following the electroporation process; and a plurality of valves connecting the plurality of chambers to the flow channel.

[0005] Generally, in one aspect, embodiments relate to a method. The method includes performing an electroporation process by generating multiple electric fields in multiple chambers using multiple electrodes, wherein the multiple chambers are configured to store multiple cells during the electroporation process. The method further includes performing a cell collection process by opening multiple valves connected to the multiple chambers; and transferring the multiple cells to an outlet using flow channels connected to the multiple valves, wherein the multiple chambers, multiple electrodes, multiple valves, outlet, and flow channels are located within an electroporation apparatus.

[0006] Other aspects of the embodiments will become apparent from the following description and the appended claims. Attached Figure Description

[0007] Figure 1 A perspective view of an electroporation apparatus according to one or more embodiments is shown.

[0008] Figure 2 A cross-section of an electroporation device according to one or more embodiments is shown.

[0009] Figure 3 A top view of a chamber according to one or more embodiments is shown.

[0010] Figure 4 A flowchart according to one or more embodiments is shown.

[0011] Figure 5 A perspective view of a seal according to one or more embodiments is shown.

[0012] Figure 6 A cross-section of a sealing cap (also known as a chamber cap) according to one or more embodiments is shown.

[0013] Figure 7 A side view of a single electroporation chamber according to one or more embodiments is shown.

[0014] Figure 8 Another side view of a single electroporation chamber according to one or more embodiments is shown.

[0015] Figure 9 Multiple electroporation chambers in an electroporation apparatus according to one or more embodiments are shown.

[0016] Figure 10 An example docking station according to one or more embodiments is shown.

[0017] Figure 11 A cross-sectional view of a seal according to one or more embodiments is shown.

[0018] Figure 12 A diagram of a valve (i.e., a chamber valve) according to one or more embodiments is shown.

[0019] Figure 13 A front view of the lever portion of a chamber valve according to one or more embodiments is shown.

[0020] Figure 14 A bottom view of an electroporation device according to one or more embodiments is shown.

[0021] Figure 15 Cross-sectional views of an example inlet pump and an example outlet pump according to one or more embodiments are shown.

[0022] Figure 16 An exploded view of an electroporation apparatus according to one or more embodiments is shown.

[0023] Figure 17 An assembly diagram of an electroporation apparatus according to one or more embodiments is shown.

[0024] Figure 18An example of a single electroporation process according to one or more embodiments is shown.

[0025] Figure 19 A flowchart illustrating the operation of an electroporation patch station according to one or more embodiments is shown. Detailed Implementation

[0026] Numerous specific details are set forth in the following detailed description of the embodiments to provide a more thorough understanding of the technology disclosed herein. However, it will be apparent to those skilled in the art that the technology disclosed herein can be practiced without these specific details or with equivalent substitutes in form and / or function.

[0027] Throughout the application, ordinal numbers (e.g., first, second, third, etc.) may be used as adjectives for elements (i.e., any noun in the application). The use of ordinal numbers is not intended to imply or create any particular ordering of elements, nor is it intended to limit any element to merely a single element, unless explicitly stated otherwise, such as through the use of the terms “before,” “after,” “single,” and other such terms. Rather, the use of ordinal numbers is to distinguish elements. As an example, the first element is distinct from the second element, and the first element may inherit from the second element in the ordering of elements (or precede the second element).

[0028] One or more embodiments relate to an electroporation apparatus and a method of using / operating the electroporation apparatus. The electroporation apparatus is capable of performing large-scale electroporation processes.

[0029] Figure 1 An electroporation device (100) according to one or more embodiments is illustrated. The electroporation device (100) may be referred to as a cassette (or dark box). The electroporation device may be sterile. The electroporation device (100) may include a housing made of plastic (e.g., polycarbonate), glass, or other materials suitable for biological and / or medical use. Figure 1 As shown, the electroporation device (100) has multiple components, including multiple openings (105), an inlet (110), an outlet (115), multiple electrodes (120), and multiple pump connectors (125). Figure 2 As further discussed and described, the electroporation apparatus (100) may additionally include pumps (e.g., diaphragm pumps; each pump having two check valves (e.g., inlet check valves and outlet check valves that allow only one-way (unidirectional) flow of liquid)) to influence fluid movement throughout the electroporation apparatus (100). Each component is discussed below.

[0030] In one or more embodiments, multiple openings (105) lead to chambers (discussed further below). Cells (along with any accompanying suspension material) can be deposited into one or more chambers via the multiple openings (105). Chemicals, drugs, and / or macromolecules (such as proteins and nucleic acids (such as various forms of DNA and RNA)) to be introduced into the cells during the electroporation process can also be deposited into the chambers via the multiple openings (105). Although Figure 1 Eight openings (and therefore eight chambers) are shown, but in other embodiments, a different number of openings (and therefore a different number of chambers) may be present. For example, in some embodiments, the cartridge may have 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25…etc., with the number of chambers increasing sequentially as needed to increase cell electroporation capability (or batch electroporation capability). In one or more embodiments, multiple chambers may share the same opening.

[0031] In one or more embodiments, each of the plurality of electrodes (120) is associated with one of the chambers. Furthermore, each of the plurality of electrodes (120) has an inner portion and an outer portion. The inner portion is inside the chamber and contacts the contents (e.g., cells) stored within the chamber. The outer portion is outside the chamber and is exposed on and / or protrudes from the surface of the electroporation device (100). The inner and / or outer portions may include elliptical (e.g., circular) surfaces. Other shapes (e.g., rectangular) are also feasible. Each of the electrodes (120) may include a base made of one metal or alloy and a coating made of the same or different metals or alloys. For example, each of the plurality of electrodes (120) may include a base made of aluminum and a gold coating. Other metals (e.g., copper, silver, etc.) may also be used instead of aluminum and / or gold, or as a complement to aluminum and / or gold. The metals and / or alloys may be selected based on chemical inertness, thus making it unlikely to chemically react with or penetrate the contents (e.g., cells) of the chamber.

[0032] In one or more embodiments, the electrodes are located on opposite surfaces of the electroporation apparatus (100). In other words, multiple electrodes (120) can be replicated on opposite surfaces. Thus, each chamber can be associated with an electrode pair (one electrode on each surface) on opposite sidewalls of that chamber. The electroporation process can be performed by applying a voltage across the electrode pair, thereby generating an electric field in the chamber associated with the electrode pair.

[0033] In one or more embodiments, the inlet (110) and outlet (115) are located at opposite ends of the electroporation apparatus (100). The inlet (110) and outlet (115) may be located on the same or different surfaces (such as the top or bottom surface) of the electroporation apparatus (100). The inlet (110) serves as an input of liquid medium during the cell collection process. For example, the liquid medium obtained at the inlet (110) can be used to flush the chamber after the electroporation process. In one or more embodiments, the inlet (110) is configured to connect to a bag (or other container) storing the liquid medium via a male Luer lock fitting (not shown). The outlet (115) serves as a collection point during the cell collection process. After the electroporated cells (in the liquid medium) and cell-free liquid medium (for flushing the chamber) have been transferred through the flow channel, the outlet (115) obtains the cells from the chamber. In one or more embodiments, the outlet (115) is configured to connect via a convex Luer lock fitting (not shown) to a bag (or other container) storing the collected cells and the collected liquid medium.

[0034] In one or more embodiments, the fluid device (e.g., a pump) may be connected to a pump connector ( Figure 1 (125) is intended for use during the cell collection process to facilitate the movement of fluids within the electroporation apparatus (100), such as the movement of cells collected after the electroporation process. The fluid apparatus and the cell collection process are discussed below.

[0035] Figure 2 A linear cross-sectional view of an electroporation device (100) according to one or more embodiments is shown. Figure 2 As shown, the electroporation apparatus (100) includes multiple chambers (205), multiple valves (210), flow channels (215), multiple side flow channels (e.g., side flow channel A (220A), side flow channel B (220B)), multiple pumps (e.g., pump A (225A), pump B (225B)), and an airflow passage (230) with vents (235). Pump A 225A and pump B 225B may be referred to as the inlet pump and the outlet pump, respectively. Each of these components is discussed below.

[0036] In one or more embodiments, the chamber (205) is configured to store cells along with chemicals, drugs, and / or macromolecules, such as proteins and nucleic acids to be introduced into the cells during the electroporation process. The chamber (205) may be formed from the housing of the electroporation device (100) and therefore may be formed from plastic (e.g., polycarbonate). In one or more embodiments, the lower portion of each chamber (205) is teardrop-shaped, as described below regarding... Figure 7The discussion continues. In other words, the lower wall of the chamber slopes inward toward the bottom of the chamber (i.e., the chamber becomes narrower). This can facilitate the emptying of the chamber (205) (discussed below). The chambers (205) can be designed to store any desired volume in each chamber, including, for example, at least 250 microliters (µL), 300 µL, 350 µL, 400 µL, 450 µL, 500 µL, 600 µL, 640 µL, 700 µL, 750 µL, 800 µL, 900 µL, 1 milliliter (mL), 2 mL, etc. Different chambers (205) can have different sizes, and different chambers (205) can store different volumes. In one or more embodiments, the chambers are designed to store a range of 300 to 640 µL (cell volume in liquid suspension) for electroporation. In one or more embodiments, the chambers are designed to store a maximum cell volume of 600 µL in liquid suspension for electroporation. In one or more embodiments, the chamber is designed to store a maximum cell volume of 640 μL in a liquid suspension for electroporation.

[0037] As described above, the electroporation device may have eight chambers (120). The combination of these eight chambers may be configured to store cells in a liquid suspension of at least 2 mL (e.g., 250 μL × 8 chambers), at least 2.4 mL (e.g., 300 μL × 8 chambers), at least 3.2 mL (e.g., 400 μL × 8 chambers), at least 4 mL (e.g., 500 μL × 8 chambers), at least 4.8 mL (e.g., 600 μL × 8 chambers), at least 5.6 mL (e.g., 700 μL × 8 chambers), or at least 6.4 mL (e.g., 800 μL × 8 chambers) for electroporation.

[0038] In one or more embodiments, a valve (210) connects a chamber (205) to a flow path (215). (See, for example, Figure 2 Each chamber may have one valve. Alternatively, multiple chambers may share a single valve. Each valve in the valves (210) may correspond to an umbrella valve, a clamp valve, a piston valve, a gate valve, a spring valve, a lever valve, etc. The valves (210) may be "off-the-shelf" (i.e., commercially available) valves of the types described above. Preferably, the selection of valves may reduce the likelihood of leakage, reduce the likelihood of blockage, and increase the number of cells collected during the cell collection process (as described below). The default position of the valves (210) is closed. Multiple valves (210) may be opened simultaneously. Alternatively, the valves (210) may be opened sequentially, such as one at a time.

[0039] In some embodiments, each chamber valve is a clamping valve and is leak-free up to at least 35 pounds per square inch (PSI) and leaks to a negative pressure of at least -10 (negative 10) PSI.

[0040] Figure 12 A diagram of a valve (1200) according to one or more embodiments is shown for both the open and closed positions of the valve. The valve (1200) may correspond to the above-described reference. Figure 2 Any of the valves (210) discussed. A valve (1200) may include a lever portion (1201) and a spring (1210). The lever portion (1201) may include a spring connector (1206) wherein the spring (1210) is attached to the lever portion (1201). The lever portion (1201) may also include a hinge (1203), a dome (1205), and a force portion (1207).

[0041] The valve (1200) is associated with a cavity in the chamber (205). In one or more embodiments, when the valve (1200) is closed, the dome (1205) shifts and compresses the rubber layer between the outlet at the bottom of the chamber and the flow channel (215). This effectively blocks the outlet at the bottom of the chamber and prevents the contents of the chamber from draining into the flow channel (215) and / or prevents liquid in the flow channel (215) from rising into the chamber. In one or more embodiments, the rubber layer is a flexible portion of the flow channel (215). When not subjected to any external force, the spring (1210) holds the valve (1200) in the closed position.

[0042] In one or more embodiments, to open the valve (1200), a force is applied to the force portion (1207) of the lever portion (1201). For example, the force can be applied by a valve actuator of the plug station (as described below). In response to the force, the lever portion (1201) rotates about the hinge (1203). This movement of the lever portion (1201) also causes the dome (1205) to move and pull out the outlet at the bottom of the chamber. Thus, when the outlet at the bottom of the chamber is pulled out, (e.g., when under the influence of a pumping force) the contents of the chamber can drain into the flow channel (215) and / or liquid in the flow channel (215) can rise into the chamber. When the force is removed from the force portion (1207), the spring (1210) returns the valve (1200) to the closed position. In other words, the spring (1210) causes the lever portion (1201) to rotate about the hinge (1203), which causes the dome (1205) to shift and compress the rubber layer, thereby effectively blocking the outlet.

[0043] Figure 13 A front view of the lever portion (1201) according to one or more embodiments is shown. Figure 13 As shown, the lever portion (1201) includes a hinge (1203), a dome (1205), and a spring connector (1206).

[0044] Figure 14A bottom view of an electroporation apparatus (100) according to one or more embodiments is shown. In this bottom view, both the flow channel (215) of the chamber (205) and the chamber outlet (e.g., chamber outlet (1405)) are visible. When the valve (1200) is closed, the dome (1205) blocks the chamber outlet (1405). As described above, this prevents the contents of the chamber from draining into the flow channel (215) and / or prevents liquid in the flow channel (215) from rising into the chamber. When the valve (1200) is open, the dome (1205) no longer blocks the chamber outlet (1405), and the contents of the chamber can drain into the flow channel (215). Similarly, if subjected to pumping force or other forces capable of moving liquid (e.g., gravity (gravity flow), increased air pressure, etc.), liquid in the flow channel (215) can rise into the chamber.

[0045] Figure 15 A cross-sectional view of an example inlet pump (225A) and an example outlet pump (225B) according to one or more embodiments is shown. The example pumps (225A, 225B) in this figure are integrated into a line with side channels (220A, 220B). As depicted herein, each pump (225A, 225B) has a flexible (e.g., silicon) diaphragm (1508) adjacent to a fluid chamber juxtaposed with a "duckbill" check valve (1506) for regulating single-pass (unidirectional) fluid flow. Each of the pumps (225A, 225B) is operated by repeatedly flattening the "dome" of the diaphragm (1508) (via a plug-in actuator) to displace the fluid. In some embodiments, each of the pumps (225A, 225B) has a normal operating flow rate of approximately 15 ml / min at 300 RPM (revolutions per minute) of the pump actuator and a “fast flow” operation of approximately 30 ml / min at 600 RPM. In some embodiments, the pump flow rate can be adjusted in increments of 50 μL. In some embodiments, each of the pumps (225A, 225B) can also function as a valve and is leak-free up to at least 35 psi (psI) and leak-free at a negative pressure of at least -10 (-10) PSI.

[0046] Figure 15 Also shown is a convex Luer lock fitting (1504) inserted into both the inlet (110) and the outlet (115). The convex Luer lock fitting (1504) is covered by a Luer cap (1502).

[0047] Return to reference Figure 2In one or more embodiments, side channels (220A, 220B) connect the channel (215) to the inlet (110) and the outlet (115). Each of the channels (220A, 220B, 215) may be a tube formed in the housing or otherwise made of plastic (e.g., polycarbonate), glass, metal, etc. During the cell collection process, the contents of the chamber (210) (e.g., a liquid suspension of cells) can be drained into the channel (215) by opening the valve (210). The liquid medium (obtained at the inlet (110)) can travel to the channel (215) via side channel A (220A) and push the drained contents (e.g., a liquid suspension of cells) from the channel (215) to the outlet (115) via side channel B (220B). Furthermore, the liquid medium can enter the chamber through an open valve (i.e., the liquid medium enters the chamber from the flow channel (215)) and collect additional cells by flushing the chamber before advancing to the outlet (115) via the side flow channel B (220B). Thus, at least one of the flow channel (215) and the side flow channel (e.g., 220B) is configured to transport electroporated cells during the cell collection process.

[0048] In one or more embodiments, one or more pumps (pump A (225A), pump B (225B)) are used to move the liquid medium and thus flush the chamber (205) and push the cells toward the outlet (115). As described above, pump A 225A and pump B 225B may be referred to as the inlet pump and the outlet pump, respectively. The number and volume of pump strokes required to flush a given chamber and push the discharged contents (i.e., the liquid suspension of cells) toward the outlet (115) depend, for example, on the distance between the given chamber and the inlet (110).

[0049] In one or more embodiments, an airflow channel (230) connects the airflow between a plurality of chambers (205) below the sealing cap (500). The airflow channel (230) is connected to the outside of the electroporation device (e.g., to maintain atmospheric pressure) via a vent or filter (235) (e.g., a microbial air filter; such as a commercially available 0.2-micron filter). After cells are deposited into the chambers (205), but before the electroporation process is performed, a seal made of, for example, silicon (or other biocompatible material) is used (regarding...). Figure 5 and Figure 6(Further discussion) Blocking (covering) the opening (105). This effectively creates a closed system. The external vent (235) or filter and the airflow channel (230) reduce or eliminate the possibility of partial vacuum formation (e.g., the air pressure in the chamber is lower than atmospheric pressure), and thus facilitate chamber discharge (exhaust channel (215)) during the cell collection process while maintaining the sterile integrity of the chamber. In one or more embodiments, pressurized air may be forced into the vent (235) and thus into the airflow channel (230) to accelerate chamber discharge during the cell collection process (wherein the amount of such pressurized air is insufficient to lift or open the sealing cap).

[0050] As described above, there are electrodes (120) associated with the chamber (205). Also as described above, the interior portion of each electrode may have an elliptical (e.g., circular) surface. Figure 2 The elliptical surface of electrode (120) is shown. In one or more embodiments, the elliptical surface is a circle with a diameter of 19.5 mm or about 19.5 mm. Other diameters and electrode shapes are also possible. In one or more embodiments, the elliptical (or circular) shape increases the conductivity across the electrode surface.

[0051] Figure 3 A representative top view of a single chamber (305) according to one or more embodiments is shown. The chamber (305) may correspond to the above-described reference. Figure 2 Any one of the chambers (205) under discussion. Chambers (305) have opposing edges (330A, 330B). As... Figure 3 As shown, the chamber (305) is associated with an electrode pair (electrode A (320A), electrode B (320B)). The two electrodes (320A, 320B) correspond to those mentioned above. Figure 1 and Figure 2 The electrodes (120) under discussion. Electrode pairs (320A, 320B) are positioned on opposite sides of a chamber (305). In one or more embodiments, the inner surfaces of the electrodes (320A, 320B) form opposite sidewalls (340A, 340B) of the chamber (305). In one or more embodiments, the inner surface of each of the electrodes (320A, 320B) is adjacent to an existing sidewall of the chamber (305). As described above, during the electroporation process, a voltage is applied across the electrodes (320A, 320B) to generate an electric field within the chamber (305). Each electrode in the electrode pair may be spaced apart from each other by a distance sufficient to reduce or eliminate arcing between the electrodes, but close enough to allow an electric field to be maintained between the electrodes. For example, the face of electrode 320A at sidewall 340A may be spaced approximately 4 millimeters (mm) from the face of electrode 320B at sidewall 340B. Other spacing distances (e.g., approximately 1 mm, 3 mm, 5 mm, 7 mm, 10 mm, etc.) are also feasible.

[0052] Figure 7 A side view of a single chamber (305) according to one or more embodiments is shown, in which no electrodes are installed. Figure 7 In one embodiment, the chamber (305) has an inverted teardrop-shaped cross-section (i.e., the chamber narrows at the bottom (or towards the bottom)). This teardrop shape facilitates the drainage of electroporated cells from the chamber (305) into the underlying flow channel (215). Although Figure 7 The embodiments shown depict an inverted teardrop-shaped cross-section, but those skilled in the art will recognize that other chamber shapes, including circular, rectangular, triangular, rhomboid, tubular, etc., may also be used.

[0053] In one or more embodiments, an outer edge (702) surrounds the edge of the chamber (305). The outer edge (702) supports one of the electrode pairs (320A, 320B); a similar outer edge exists on the opposite side of the chamber (305) for supporting the other of the electrode pairs (320A, 320B). Figure 8 The image shows the electrode (320A) mounted on it. Figure 7 Side view.

[0054] return Figure 7 The edge surfaces (330A, 330B) of the chamber (305) (which may include multiple bottom surfaces) can be formed by the housing of the electroporation device (100). Once the electrodes (320A, 320B) are inserted into the outer edge (702), the sidewalls of the chamber (305) can be formed. Figure 3 (340A, 340B).

[0055] Figure 9 A plurality of chambers (305) arranged adjacent to each other according to one or more embodiments are shown. In one or more embodiments, the electrodes (320) are separated by a distance 902 to eliminate the interaction between adjacent electrodes (320).

[0056] As mentioned above Figure 2 As discussed, in one or more embodiments, once the necessary materials (e.g., a liquid suspension of cells and electroporation materials (e.g., nucleic acids)) have been added to the chamber (205), the opening (105) at the top of the chamber (205) can be sealed with a seal to create a self-contained, biosafe device. Figure 5 An example seal 500 is shown that can be used to seal multiple openings (105). Figure 5 Includes multiple sealing caps (502A-502H), each sealing cap (502) corresponding to one of the openings (105). Although Figure 5A plurality of sealing caps (502) connected together via bridging portions (504A-504G) are shown, but those skilled in the art who benefit from this detailed description will recognize that each sealing cap (502) may be applied individually to each of the openings (105), or a smaller set of sealing caps (502) may be connected together to seal a subset of the openings (105).

[0057] Each sealing cap (502) includes a top (506) and a bottom protrusion (508). Figure 6 A cross-section of a sealing cap (502) according to one or more embodiments is shown. The top (506) of the sealing cap (502) is configured to seal the top of a corresponding chamber (205). A bottom protrusion (508) is configured to extend into a corresponding opening (105) of the corresponding chamber (205) to ensure a tight fit between the opening (105) and the sealing cap (502). In one or more embodiments, although the bottom protrusion (508) partially extends into the corresponding opening (105), sufficient space is left between the bottom lip (508) and the corresponding chamber (205) to allow airflow between the chamber (205) and the airflow passage (230). Similarly, when the seal (500) is installed across multiple chambers (205), sufficient space is left below the bridging portion (504) to allow airflow through the airflow passage (230). As the airflow channel (230) is vented through the air filter or vent (235), the seal (500) creates a biologically closed containment system that still allows the air pressure within the system to be maintained.

[0058] Figure 11 A cross-sectional view is shown of a seal (500) inserted into an opening (105) of a corresponding chamber (205) according to one or more embodiments. (See from...) Figure 11 As can be seen, the sealing cap of the seal (500) presses into and interferes with the conical chamber wall at the opening (105) to create a seal. Double-headed arrows indicate the spacing between the internal electrode faces. A vent (or microbial air filter) is indicated by a circle on the left side of the chamber cap (500).

[0059] Figure 16 An exploded view of the electroporation device (100) (which (as described above) may also be referred to as a box) is shown. Figure 16 In the diagram, prior to the assembly of the electroporation device (100), multiple electrodes (120), an inlet (110), an outlet (115), a convex Luer lock fitting (1504), a Luer cap (1502), and a seal (500) are shown. Figure 16 Pump housing A (1605A) and pump housing B (1605) configured to store components of inlet pump (225A) and outlet pump (225B) respectively are also shown.

[0060] Figure 17 An assembly diagram of the electroporation device (100) (which (as described above) may also be referred to as a box) is shown. Figure 17 Multiple electrodes 120, pump housing A (1605A), pump housing B (1605B), multiple chamber valves (210) (each valve having a spring (1210) and a lever (1201)), seals (500), convex Luer lock fittings (1504) and Luer caps (1502) are shown.

[0061] Figure 4 A flowchart is shown according to one or more embodiments of the present invention. Figure 4 The flowchart depicts the process for using / operating the electroporation device (100) described above. In one or more embodiments, details may be omitted, repeated, and / or used in conjunction with... Figure 4 The different sequences shown are executed sequentially. Figure 4 One or more of the steps shown. Therefore, the scope of the invention should not be considered limited to. Figure 4 The specific arrangement of the steps shown.

[0062] In step 407, cells and chemicals, drugs, and / or macromolecules (such as proteins and nucleic acids) to be introduced into the cells are loaded into the chambers (205) of the electroporation device (100). This loading may occur via, for example, an opening (105). The opening (105) can then be sealed using seals (such as a seal (500) and / or a sealing cap (502)). Although the electroporation device (100) has multiple chambers, some chambers may not be used (i.e., a liquid suspension of cells may not deposit in some chambers).

[0063] In step 409, the electroporation device (100) is loaded into the plug station. Figure 10 An example plug-in station (1000) according to one or more embodiments is shown. The plug-in station (1000) includes a socket (1002), a valve actuator (1004), electrical contacts (1006), and a pump actuator (1008). The socket (1002) is sized and shaped to receive an electroporation device (100) and hold the electroporation device (100) in a fixed and upright position. The valve actuator (1004) is configured to engage a valve (210) on the electroporation device (100). For example, if the valve (210) on the electroporation device (100) is a spring-loaded valve, the valve actuator (1004) would include components that apply force (e.g., press) to the valve (210) to open the valve (210). In one or more embodiments, each valve actuator (1004) has a one-to-one correspondence with a valve (210), such that each valve (210) can be individually controlled by the corresponding valve actuator (1004).

[0064] The electrical contacts (1006) of the plug-in station (1000) engage with the electrodes (120) of the electroporation device (100). For example... Figure 10 As shown, the electrical contacts (1006) can be linearly aligned along the length of the socket (1002). Furthermore, the electrical contacts (1006) are positioned on opposite sides of the socket (1002); for viewing purposes, Figure 10 Only one side (1002) is depicted. The electrical contact (1006) can be, for example, a high-voltage contact. The pump actuator (1008) of the plug-in station (1000) engages with the fluid components (such as pumps (pump A (225A), pump B (225B))) of the electroporation device (100). Each of the valve actuator (1004), the electrical contact (1006), and the pump actuator (1008) can be controlled by one or more control panels or devices (not shown) operatively linked to the plug-in station.

[0065] return Figure 4 At step 409, due to the loading of the electroporation device (100) into the plug station, the exterior of each of the electrodes (120) comes into contact with one or more circuits of the plug station (such as electrical contacts (1006) of the plug station (1000)). Thus, after the electroporation device (100) is loaded into the plug station, the electrodes (120) become elements of one or more circuits. Furthermore, due to the loading of the electroporation device (100) into the plug station, one or more pumps (225A, 225B) and valves (210) or valve lever portions (1201) can be operatively contacted with actuators of the plug station. A bag (or other container) containing a liquid medium can be attached to the inlet (110), and a collection bag (or other container) can be attached to the outlet (115) of the electroporation device (100).

[0066] In step 412, an electric field can be generated in one or more of the chambers (205) using electrodes (120). For example, the plug-in station can use circuitry controlled by software (e.g., via a linked computer device) to apply one or more voltage pulses to the electrodes (120) to generate an electric field. Electric fields can be generated simultaneously in all chambers (205). Alternatively, an electric field can be generated for one chamber (205) at a time, or for a subset of chambers (205) at a time. These applied electric fields increase cell membrane permeability and thus allow the introduction of chemicals, drugs, and / or macromolecules (such as proteins and nucleic acids) into the cell.

[0067] In step 414, the valve (210) of the electroporation device (100) is opened. For example, the valve actuator (1004) of the plug station (1000) can open the valve (210) of the plugged electroporation device (100). The plug station can open all valves (210) simultaneously. Alternatively, the plug station can open one valve (210) at a time, or a subset of valves (210) at a time. Depending on the type of valve, the actuator may need to manipulate a piston, lever, spring, etc., to open the valve (210). In other words, the valve (210) can be operated using spring movement, lever movement, piston movement, etc. Opening one of the valves (210) causes the contents of the chamber connected to the valve to drain into the flow channel (215). Such discharge may result from one or more of the following: hydraulic pressure generated by the actuation of one or more pumps; gravity (depending on the orientation of the valve (210) relative to the chamber (205); pressure difference between the chamber (205) and the flow channel (215); increased air pressure; capillary effect; and so on. In one or more embodiments, a venting airflow channel (230) extending below the opening (105) and between the chambers (205) can aid the discharge process by preventing the creation of a partial vacuum. In one embodiment, pressurized air may be forced into an air filter or vent (235) connecting the airflow channel (230) to the outside of the electroporation device (100) to accelerate the discharge process.

[0068] In step 416, liquid medium is pumped from the inlet (110) into the chamber (205) of the electroporation apparatus (100), and the electroporated cells are collected at the outlet (115). For example, the pump actuator (1008) of the plug station (1000) can operate one or more pumps (225A, 225B) to pump liquid medium from a bag (or other container) attached to the inlet (110) into the electroporation apparatus (100). Operating the pumps (225A, 225B) causes the liquid medium to travel through various channels (220A, 215, 220B) and to deliver the liquid suspension of discharged cells in the flow channel (215) to the outlet (115) and into the collection bag (or other container) connected to the outlet (115). Operating the pumps (225A, 225B) also forces the liquid medium from the flow channel (215) (via an open valve) into the chamber (205) so that any residual / residual cells remaining in the chamber (205) are flushed before the liquid medium delivers cells through the flow channel (215) to the outlet (115). The chambers (205) can be flushed simultaneously. Alternatively, one chamber (205) can be flushed at a time, or a subset of chambers (205) can be flushed together. Furthermore, each chamber can be flushed immediately after discharge.

[0069] In one or more embodiments, step 412 corresponds to the electroporation process, while steps 414 and 416 correspond to the cell collection process performed after the electroporation process.

[0070] Figure 19 A flowchart according to one or more embodiments is shown. Figure 19 The flowchart depicts the process for using / operating the above reference. Figure 10 The process of the aforementioned plug-in station (1000). In one or more embodiments, it may be omitted, repeated, and / or used in conjunction with... Figure 19 The different sequences shown are executed sequentially. Figure 19 One or more steps are shown. Therefore, the scope of the invention should not be considered limited to... Figure 19 The specific arrangement of the steps shown. Figure 19 The process described in the middle and Figure 4 The process described in the text is related (as mentioned above).

[0071] In step 1907, the electroporation device (100) (in one embodiment, pre-loaded with cells in a liquid suspension) is secured in a socket (1002) of a connector (1000). The socket (1002) includes an opening for inserting the electroporation device (100) and securing it in an upright position. After securing the electroporation device (100) into the socket, the electrical contacts (1006) of the connector (1000) are brought into contact with the electrodes (120) of the electroporation device (100). As described above, the electrical contacts (1006) are positioned on opposite sides of the socket (1002).

[0072] Similarly, after fixing the electroporation device (100), the valve actuator (1004) of the plug station (1000) can engage with the valve (210) of the electroporation device (100), and the pump actuator (1008) of the plug station (1000) can engage with the pump (225A, 225B) of the electroporation device (100).

[0073] One or more chambers (205) of the electroporation device (100) may be filled (via deposition of a liquid suspension) with cells and chemicals, drugs, and / or macromolecules (such as proteins and nucleic acids) to be introduced before the electroporation device (100) is secured in the socket (1002). Furthermore, a seal (500) may be positioned at the opening (105) of the electroporation device (100) before the electroporation device (100) is secured in the socket (1002). Before or after the electroporation device (100) is secured in the socket (1002), a bag (or other container) containing a liquid medium may be attached (via a convex Luer lock fitting 1504) to the inlet (110) of the electroporation device (100), and a collection bag (or other container) may be attached (via a convex Luer lock fitting 1504) to the outlet (115) of the electroporation device (100).

[0074] At step 1909, the electroporation plug-in station (1000) uses electrical contacts (1006) to generate an electric field between electrode pairs (120) in the chambers (205) of the electroporation device (100). The electrical contacts (1006) are elements in the circuitry of the plug-in station (1000). The electric field can be generated by driving the electrical contacts (1006) with one or more signals using a pulse generator. Electric fields can be generated simultaneously in all chambers (205). Alternatively, an electric field can be generated for one chamber (205) at a time, or for a subset of chambers (205) at a time. These applied electric fields increase the permeability of cell membranes and thus allow the introduction of chemicals, drugs, and / or macromolecules such as proteins and nucleic acids into the cell.

[0075] At step 1912, the valve actuator (1004) of the plug station (1000) is operated to open the valve (210) of the docked electroporation device (100). The plug station (1000) may open all valves (210) simultaneously. Alternatively, the plug station (1000) may open one valve (210) at a time, or a subset of valves (210) at a time. Depending on the type of valve, the actuator may need to manipulate pistons, levers, springs, etc., to open the valve (210). Opening one of the valves (210) allows the contents of the chamber connected to the valve to drain into the flow path (215) of the electroporation device (100).

[0076] At step 1914, the pump actuator (1008) of the plug-in station (1000) is operated to activate the pumps (225A, 225B). This may include repeatedly flattening the diaphragm (1508) of each pump (225A, 225B). As a result, liquid media are pumped from a bag (or other container) attached to the inlet (110) into the electroporation device (100). Specifically, the pump actuator (1008) is operated to cause the pumps (225A, 225B) to pump the liquid media through various channels (220A, 215, 220B) and to deliver a liquid suspension of discharged cells in the flow channel (215) to the outlet (115) and into a collection bag (or other container) attached to the outlet (115). Operating the pumps (225A, 225B) also forces the liquid medium from the flow channel (215) (via an open valve) into the chamber (205) so that any residual / residual cells remaining in the chamber (205) are flushed before the liquid medium is transferred to the outlet (115) via the flow channel (215). The chambers (205) can be flushed simultaneously. Alternatively, one chamber (205) can be flushed at a time, or a subset of chambers (205) can be flushed together. Furthermore, each chamber can be flushed immediately after discharge.

[0077] In one or more embodiments, step 1909 corresponds to the electroporation process, while steps 1912 and 1914 correspond to the cell collection process performed after the electroporation process.

[0078] In one or more embodiments, the electroporation device (100) is sterilized. In one or more embodiments, the electroporation device (100) is sterilized by exposure to gamma radiation at a dose of 50 kilogray (kGy) or greater. In one or more embodiments, the electroporation device (100) is sterilized by exposure to gamma radiation at a dose of 50-70 kilogray (kGy). In one or more embodiments, the electroporation device (100) is fully functional after the sterilization process. In one or more embodiments, the electroporation device (100) is fully functional after exposure to gamma radiation at a dose of 50-70 kilogray (kGy).

[0079] In one or more embodiments, the electroporation device (100) is for single use. In one or more other embodiments, the electroporation device (100) can be reused. In other words, a single electroporation device can be reused multiple times. Figure 4 and / or Figure 19 The process described in the text.

[0080] Traditional electroporation systems require the use of multiple cuvettes to electroporate large numbers of cells. Furthermore, even though a biosafety cabinet (BSC) can be used to provide aseptic conditions in such processes (i.e., transferring cells to multiple cuvettes), handling multiple cuvettes inherently increases the chance of introducing microbial contamination (i.e., loss of aseptic conditions). Additionally, such multiple treatments inherently increase processing time and introduce unavoidable variations in conditions and / or process consistency.

[0081] As a significant improvement over previous systems, the electroporation device (100) and the insertion station (1000) can be used to electroporate a large number of cells in a single electroporation process (i.e., in a single electroporation “run”).

[0082] In one or more embodiments, the electroporation device (100) and the insertion station (1000) can be used to perform electroporation on a single electroporation process (i.e., in a single "run"), for example, but not limited to, at least 1 × 10 8 1 cell, at least 2 × 10 8 1 cell, at least 3 × 10 8 1 cell, at least 4 × 10 8 1 cell, at least 5 × 10 8 1 cell, at least 6 × 10 8 1 cell, at least 7 × 10 8 1 cell, at least 8 × 10 8 1 cell, at least 9 × 10 8 10 cells, at least 1×10 9 10 cells, at least 2 × 10 9 1 cell, at least 3 × 10 9 1 cell, at least 4 × 10 9 1 cell, at least 5 × 10 9 1 cell, at least 6 × 10 9 1 cell, at least 7 × 10 9 1 cell, at least 8 × 10 9 1 cell, at least 9 × 10 9 10 cells, at least 1×10 10 1 cell, at least 2 × 10 10 1 cell, at least 3 × 10 10 1 cell, at least 4 × 10 10 1 cell, at least 5 × 10 10 1 cell, at least 6 × 10 10 1 cell, at least 7 × 10 10 1 cell, at least 8 × 10 10 1 cell, at least 9 × 10 10 1 × 10 cells, at least 1 × 10 111 cell, at least 2 × 10 11 1 cell, at least 3 × 10 11 1 cell, at least 4 × 10 11 1 cell, at least 5 × 10 11 1 cell, at least 6 × 10 11 1 cell, at least 7 × 10 11 1 cell, at least 8 × 10 11 1 cell, at least 9 × 10 11 10 cells, at least 1×10 12 1 cell, at least 2 × 10 12 1 cell, at least 3 × 10 12 1 cell, at least 4 × 10 12 1 cell, at least 5 × 10 12 1 cell, at least 6 × 10 12 1 cell, at least 7 × 10 12 1 cell, at least 8 × 10 12 10 cells, and at least 9 × 10 12 Electroporation was performed on individual cells.

[0083] In one or more embodiments, the electroporation device (100) and the insertion station (1000) can be used for electroporation of any type of eukaryotic or prokaryotic cells (e.g., but not limited to non-adherent cells, such as immune cells, NK cells, T cells, etc.).

[0084] Figure 18An example of a “closed” (i.e., sterile or disinfected) configuration of the electroporation apparatus components used in a single electroporation process (electroporation “run”) is shown. The run may include one or more of the following steps: under sterile conditions (e.g., in a biosafety / biosafety cabinet), a container (such as an input medium bag (1805)) is connected to an inlet (such as via an inlet disinfection tube (1815)), and another container (such as an output cell culture bag (1810); for cell collection after electroporation) is connected to an outlet (such as via an outlet disinfection tube (1820)). The electroporation apparatus or cassette (100) (now a closed system) is then placed in a plug-in station or “nest” (1000), and the remainder of the electroporation process may be controlled by a computer (e.g., a laptop or tablet) operatively linked to the “nest” along with an electroporation pulse generator (for delivering (multiple) electrical signals). One or more electric fields are generated between electrode pairs within the chamber. Electroporated cells are collected via outlet (115) (e.g., via pumping cell culture medium through the electroporation device or cassette (100)) into a collection container (such as an output cell culture bag (1810)); this collection container may be pre-filled with a certain amount of culture medium. After electroporation is complete, the output cell culture bag (1810) can be aseptically removed from the electroporation device / cassette and placed in an incubator (e.g., via sealing / closing the connection between the output cell culture bag (1810) and outlet (115) using a tubular heat sealer).

[0085] The time required to perform the entire electroporation process using one or more of the disclosed embodiments is significantly less than that required by systems that require the use of multiple individual cuvettes. Therefore, the examples of the electroporation apparatus or cartridge (100) described herein can be used to automate electroporation in a closed manner, thereby delivering higher yields of transfected cells (e.g., transfected immune cells / T cells) more efficiently and consistently than other available systems. Thus, the electroporation apparatus (100) described herein enables highly automated electroporation of large numbers of cells in a closed system (thus providing the ability to rapidly and efficiently generate large numbers of transfected cells in a sterile and / or cGMP manufacturing environment).

[0086] The containers or bags (1805, 1810) used in the electroporation process can be, for example, but not limited to, cell culture bags made of fluorinated ethylene propylene (FEP) material, to provide high permeability to oxygen and carbon dioxide while maintaining impermeability to water for improved culture and expansion.

[0087] Components of the electroporation device or cartridge (100) may include gold-plated electrodes. Gold is chosen because of its biocompatibility and good electrical properties. The electroporation device or cartridge (100) can be assembled in a controlled cleanroom environment. The electroporation device or cartridge (100) can be cleaned and sterilized by gamma irradiation prior to distribution and / or use.

[0088] As described above, the electroporation box described herein can be used within a system that also includes a computer (e.g., including a laptop or tablet), an electrical pulse generator, and a plug-in station or "nest" (1000) to allow for the fixation (e.g., holding) and automated manipulation of the electroporation box processes (e.g., applying multiple electric fields to cells within the electroporation chamber, pumping media and cells through the box (i.e., channels and chambers), opening and closing box valves (210)). In this type of system, the computer (or laptop / tablet) acts as the user interface and is operatively connected to control the electrical pulse generator. The generator provides electroporation pulses via contacts with the box electrodes, through connections in the nest. Thus, the plug-in station or "nest" (1000) holds the box and provides both mechanical and electrical contact with the box.

[0089] The example box may include eight chambers and a lid for covering and sealing the chambers after filling with cell suspension material (e.g., cells, media, nucleic acids, proteins, small molecules). The box may have two fittings (such as Luer-type fittings) (1502, 1504) to allow aseptic attachment of an input medium bag (1805) and an output cell culture bag (1810) to a biosafety cabinet. The input medium bag (1805) is filled with an appropriate amount of recycled media and attached by the user to the input fitting on the box in the biosafety cabinet (before electroporation). The output cell culture bag (1810) may be filled with a certain volume of recycled media and attached by the user to the output fitting on the box in the biosafety cabinet (again, before electroporation).

[0090] Each chamber (205) can be normally closed to prevent sample from draining into the manifold channel before electroporation. These valves (210) can be opened when actuated by the plug station or "nest" (1000). Directly below the inlet (110) and outlet (115) and fittings (1504) are diaphragm pumps (225A, 225B). The motor in the plug station or "nest" (1000) can pump fluid via a check valve built into the pump stack (i.e., inside). This system configuration ensures that the culture medium flows only unidirectionally through the chambers, manifold, and to the output cell culture bag (1810). The diaphragm pumps (225A, 225B) can also function as valves when closed.

[0091] For electroporation, the user can aseptically transfer the cell / nucleic acid mixture into the chamber (205) of the box and cover the box in a biosafety cabinet. The valve (210) in the box can remain closed until the actuator in the nest is opened. Each chamber (205) can be electroporated, then drained (by opening the valve (210)) and the diaphragm pumps (225A, 225B) actuated until the sample reaches the output cell culture bag (1810). This process can be repeated until all chambers (205) have been electroporated, drained, and pumped to the output cell culture bag (1810). After electroporation, the recovered cell culture medium from the input medium bag (1805) can be pumped through the box to flush the chambers (205) and the box channels (215, 220A, 220B). Once the flushing cycle is complete, the output cell culture bag (1810) can be aseptically removed from the box via a heat-sealed outlet sterilization tube (1820) and placed in a cell culture incubator.

[0092] In summary, the electroporation device (100) described herein represents a significant improvement in the large-scale electroporation of cells (e.g., immune cells / T cells) and the production of genetically engineered cell products. The electroporation cassette, within a closed system, electroporates large numbers of cells in a short time with minimal manual operation (i.e., in a largely automated manner), thereby significantly reducing the likelihood of microbial contamination and enhancing the consistency of cell products.

[0093] The embodiments and examples described herein are intended to best explain various embodiments and their particular applications, thereby enabling those skilled in the art to make and use these embodiments. However, those skilled in the art will recognize that the foregoing descriptions and examples are for illustrative and exemplary purposes only. The descriptions set forth are not intended to be exhaustive or limited to the precise forms disclosed.

[0094] While many embodiments have been described, those skilled in the art who benefit from this disclosure will understand that other embodiments can be devised without departing from the scope. Therefore, the scope of the invention should be defined only by the appended claims.

Claims

1. A system for electroporation of a sample, comprising: (a) An electroporation apparatus, comprising: (i) Multiple chambers, each configured to store multiple cells during the electroporation process; (ii) A plurality of electrodes configured to generate an electric field in each of the plurality of chambers during the electroporation process; (iii) a flow channel configured to transfer cells from the plurality of chambers during a cell collection process following electroporation; and (iv) A plurality of valves connecting the plurality of chambers to the flow path; and (b) Electroporation connection station, including: (i) a socket configured to secure the electroporation device in an upright position; (ii) a plurality of electrical contacts arranged for alignment with corresponding electrodes on the electroporation device when the device is secured in the socket; and (iii) A plurality of valve actuators arranged to engage with corresponding valves in the electroporation device when the device is fixed in the socket.

2. The system of claim 1, wherein, The electroporation device further includes: (a) Entrance; (b) Exports; and (c) A plurality of side channels connecting the inlet and the outlet to the channel.

3. The system of claim 2, wherein, The electroporation device further includes: A pump configured to pump a liquid medium from the flow channel into at least one of the plurality of chambers during the collection process; wherein the liquid medium is introduced at the inlet.

4. The system of claim 1, wherein, The electroporation device further includes: (a) A surface including a plurality of openings, each opening leading to one of the plurality of chambers; and (b) An airflow passage below the plurality of openings and connecting the airflow between the plurality of chambers.

5. The system of claim 4, wherein, The electroporation device further includes: A vent or air filter connects the airflow passage to the outside of the electroporation device.

6. The system as described in claim 4, characterized in that, The electroporation device further includes a seal configured to cover the plurality of openings.

7. The system as described in claim 1, characterized in that, Each of the plurality of chambers is shaped to narrow toward its associated valve.

8. The system as described in claim 1, characterized in that... The plurality of electrodes includes electrode pairs positioned on opposite sides of the chambers within the plurality of chambers.

9. The system as described in claim 8, characterized in that, Each electrode of the electrode pair includes: (a) The interior portion of the chamber; and (b) The outer portion outside the chamber; and Each electrode pair is configured to be connected to a circuit.

10. The system as described in claim 9, characterized in that, The inner portion of the electrode pair within the chamber has an elliptical surface and includes a gold coating.

11. The system as claimed in claim 1, characterized in that, Each of the plurality of chambers is configured to store a volume of at least 250 microliters.

12. The system as claimed in claim 1, characterized in that, Each of the plurality of chambers is configured to store a volume of at least 500 microliters.

13. The system as described in claim 3, characterized in that, The pump further includes a valve that allows only unidirectional fluid flow.

14. The system as claimed in claim 1, characterized in that, Each of the plurality of valves corresponds to a specific chamber among the plurality of chambers.

15. The system as described in claim 14, characterized in that, Each valve is a pinch valve.

16. The system as claimed in claim 1, characterized in that, The multiple chambers are configured in combination to store cells in a liquid suspension of at least 2 mL for electroporation.

17. A method of performing electroporation using the system of claim 1, the method comprising: (a) Depositing cells into an opening of at least one of the plurality of chambers in the electroporation apparatus; (b) Apply a seal to the opening; (c) Insert the electroporation device into the electroporation socket; (d) Electroporation is performed by generating an electric field in the cavity using multiple electrodes; as well as (e) Electroporation cells are collected by opening the valve connected to the chamber, allowing the cells to be transported to the outlet through the flow channel.

18. The method of claim 17, wherein the collection of the electroporation cells further comprises pumping the liquid medium obtained at the inlet through the flow channel into at least one of the plurality of chambers.

19. The method of claim 17, wherein the pressure within the plurality of chambers is maintained via a vent or air filter connected to an airflow passage extending between the plurality of chambers.

20. The method as described in claim 17, characterized in that, At least 1 x 10 8 cells are electroporated.

21. An electroporation socket, comprising: (a) A socket configured to secure the electroporation device in an upright position; (b) A plurality of electrical contacts arranged for alignment with corresponding plurality of electrodes on the electroporation device when fixed in the socket; as well as (c) A plurality of valve actuators, each valve actuator being arranged to engage with a corresponding valve among a plurality of valves in the electroporation device when the device is fixed in the socket, wherein each of the plurality of valves corresponds to a corresponding chamber among a plurality of chambers in the electroporation device.

22. The electroporation patch panel of claim 21, further comprising: A pump actuator, the pump brake being arranged to engage with the pump of the electroporation device when the device is fixed in the socket.

23. The electroporation patch panel as described in claim 22, further comprising: A circuit configured to generate an electric field using the electrical contacts.

24. The electroporation patch panel of claim 21, further comprising: (d) A container connected to the inlet of the electroporation device when the device is fixed in the socket; as well as (e) A container connected to the outlet of the electroporation device when the device is fixed in the socket.

25. The electroporation patch panel as described in claim 21, characterized in that, The socket includes an opening, and the electrical contacts are linearly aligned along the length of the socket.

26. The electroporation patch panel as described in claim 21, characterized in that, The plurality of electrical contacts are positioned on opposite sides of the socket.

27. The electroporation patch panel as described in claim 21, characterized in that, Each of the plurality of valves is a spring-loaded valve, and each of the plurality of valve actuators is capable of opening the valve by applying force to it.

28. The electroporation patch panel as described in claim 27, characterized in that, The plurality of valve actuators are configured to open all of the plurality of valves simultaneously.

29. The electroporation patch panel as described in claim 27, characterized in that, The plurality of valve actuators are configured to open each of the plurality of valves in sequence.

30. A method comprising: (a) In the socket of the electroporation station as described in claim 21, the electroporation device is fixed in an upright position; (b) During the electroporation process, an electric field is generated using multiple electrical contacts of the electroporation terminal; (c) During the cell collection process, operating one or more of the plurality of valve actuators of the electroporation insertion station.

31. The method of claim 30, further comprising: During the cell collection, a liquid medium is pumped through the plurality of valves by operating the pump actuator of the electroporation patch station.

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