System for automated preparation of regenerative epithelial suspensions and related methods of use

Abstract

One or more systems for automated preparation of regenerative epithelial suspensions include a base unit, a cartridge assembly including a cup, and a tissue dispersion device. The system may also include associated tools. The system executes one or more sequences in which a first motor operates a pestle against a tissue sample disposed along a screen in the cup while the tissue sample is sequentially contacted with an enzyme solution, a first buffer solution, and a second buffer solution. For transitions between fluid contacting phases or steps, the tissue dispersion device engages the cup and raises it to an elevated position within the processing area of ​​the cartridge while the base plate motor rotates and aligns the appropriate well under the cup for each processing step. Once the sequence is complete, a user may withdraw the regenerative epithelial suspension through an opening in the cartridge top cover.

Description

[Technical Field]

[0001] Related Applications This application claims priority to U.S. Provisional Patent Application No. 63 / 435,446, filed December 27, 2022, and U.S. Provisional Patent Application No. 63 / 533,586, filed August 18, 2023, the entire contents of both of which are incorporated herein by reference.

[0002] The present invention relates generally to the field of regenerative medicine. [Background technology]

[0003] In the field of regenerative medicine, it is generally known that manually prepared regenerative epithelial suspensions, such as those generally disclosed in U.S. Patent No. 9,029,140 and U.S. Patent Application No. 17 / 690,941, can aid in tissue repair, and the entire contents of both disclosures are incorporated herein by reference. While the exact process that such suspensions assist or enhance is not fully understood, the beneficial effects of regenerative epithelial suspensions in such processes have been well documented. It has also been well documented that the process of manually producing regenerative epithelial suspensions without automation is laborious and subject to user error and variability.

[0004] Other disclosures, such as U.S. Patent Nos. 11,124,752, 8,162,247, 10,801,001, 8,286,899, and even 10,857,544, disclose efforts related to automating tissue processing and producing regenerative epithelial suspensions. While each of these disclosures has merit, none of them fully address both the challenges of automating and improving the production of regenerative epithelial suspensions.

[0005] Thus, on the one hand, it is clear that regenerative epithelial suspensions and suspensions containing cell-derived biomolecules are important elements in tissue regeneration, and on the other hand, it is equally clear that the current state of automation in the art requires improvement.

[0006] Therefore, there is a need for an improved system for the automated preparation of regenerative epithelial suspensions and related methods of use. Summary of the Invention

[0007] According to a first aspect, there is provided a system for automated preparation of a regenerative epithelial suspension, which may comprise a base unit, a cartridge, and a pestle. In some embodiments, the system may further comprise a set of associated tools.

[0008] In some embodiments, the base unit may include: (a) a housing including a tissue processing area and a heating element; (b) a cartridge including: (i) a cartridge top cover having a cartridge top surface; (ii) at least one well plate below the cartridge top cover and including at least one well; (iii) a mortar cup (or cup); (iv) a raised processing opening disposed on the cartridge top surface and configured to receive the mortar cup (or cup); and (v) a mortar screen (or screen) disposed laterally within the mortar cup (or cup); and (c) a pestle including a pestle head having at least one dispersing surface.

[0009] In some embodiments, a system for automated preparation of a regenerative epithelial suspension may include a base unit. In some embodiments, the base unit may include a housing having an upper housing portion, a central housing portion, and a lower housing portion. The tissue processing area may be distributed between a lower surface of the upper housing portion, a front surface of the central housing portion, and an upper surface of the lower housing portion.

[0010] In some embodiments, a cover may be rotatably connected to the upper housing and configured to enclose the entire tissue processing area. At least one upper motor may be located within the upper housing. A power source may be coupled to the upper motor.

[0011] The base unit may further include a pestle shaft (or shaft). The upper motor may be configured to operate at least the pestle shaft (or shaft), and the pestle shaft (or shaft) may extend into the tissue processing area. At least one lower motor may be located within the lower housing portion, and the at least one lower motor may be configured to rotate the well plate. A heating element may be disposed on an upper surface of the lower housing portion. A power source may be electronically coupled to the at least one upper motor and the lower motor.

[0012] In one set of embodiments, the cartridge may include a cartridge bottom surface and at least one cartridge side surface. The cartridge top surface may include at least one (e.g., at least three) top openings (e.g., cartridge top cover openings).

[0013] Below the cartridge top surface may be a well plate, the well plate comprising at least one well (e.g., at least three wells), each of the at least three wells may comprise a well opening, and the well plate may be configured such that it can be rotated to align one or more of the well openings with one or more of the top surface openings.

[0014] One or more raised processing elements may be disposed along an inner surface of at least one well (e.g., a distal bottom surface of at least one well). A raised processing opening may be disposed on the cartridge top surface, and the raised processing opening may be configured to receive a mortar cup (or cup). A mortar screen (or screen) may be disposed laterally within the mortar cup (or cup). The mortar cup (or cup) may include at least one tab.

[0015] The pestle may include a pestle top cap, which may include an opening configured to receive the pestle shaft (or shaft). The pestle may include a pestle spring and / or a pestle spring cap configured to receive the pestle spring. The pestle may further include a pestle bottom cap and / or a pestle body, which may be configured to receive the pestle bottom cap.

[0016] The pestle may include a pestle head having at least one dispersing surface, the pestle head may be configured to receive the pestle body. The pestle head may have at least one raised surface element disposed along the at least one dispersing surface. The pestle body may be configured to nest within the pestle head.

[0017] In some embodiments, the system may further include a set of tools including at least one syringe (e.g., two syringes). The set of tools may further include at least one spray nozzle. In some embodiments, the system may further include at least one vial of enzyme solution and / or at least one vial of buffer solution.

[0018] In some embodiments, the raised surface element may comprise a plurality of pips disposed along at least one dispersing surface. In some embodiments, at least one raised side element may be disposed along the surface of the pestle head.

[0019] In some embodiments, the dispersive surface may comprise a distal end and at least one side surface. The distal end may comprise a curved surface, and the curved surface may comprise a plurality of protrusions. The curved surface may comprise raised helical elements.

[0020] The distal end may comprise a conical surface, which may comprise a plurality of protrusions. The conical surface may comprise a raised helical element.

[0021] In some embodiments, the distal end of the dispersive surface can include a mating end, which can include an outer surface and a terminal surface. The terminal surface can include a plurality of protrusions disposed along the terminal surface. The terminal surface can also include a raised helical element disposed along the terminal surface.

[0022] In some embodiments, the mortar cup (or cup) may have an internal ridge disposed along its inner surface, and the internal ridge may be configured to receive a raised side element such that the pestle head and the mortar cup (or cup) may be releasably engaged. In some embodiments, when the pestle head and the mortar cup (or cup) are releasably engaged (such as, but not limited to, by contact of one or more raised side elements with the inner surface of the mortar cup (or cup)), the pestle may be able to lift, rotate, or otherwise manipulate the mortar cup (or cup).

[0023] In some embodiments, at least one upper motor may be configured to actuate the pestle shaft (or shaft) and thus the pestle to move the pestle head back and forth along the Y-axis (which may generate a normal force), rotate (which may generate a rotational force), or both vertically and rotate (which may generate a grinding force). In some embodiments, actuation of the pestle shaft (or shaft) may cause the pestle head, and in some embodiments, the pestle head having at least one raised surface element disposed on its distal surface, to apply a normal force, a rotational force, or a grinding force, or a combination of two or more such forces, to the tissue sample. In some embodiments, the tissue sample may be disposed along a mortar screen (or screen) within a mortar cup (or cup). In some embodiments, the mortar cup (or cup) may be disposed within a well such that the mortar screen (or screen) comes into contact with one or more raised treatment elements, which may be disposed along the interior of the well. In such embodiments, it is believed that the additional opposing pressure provided by the multiple raised processing elements may enhance the dispersion effect of the normal, rotational, or grinding force, or a combination of two or more such forces, applied to the tissue sample by actuation of the pestle.

[0024] In some embodiments, the mortar screen (or screen) may be configured to separate particulates greater than 10 microns in size. In some embodiments, the mortar screen (or screen) may be configured to separate particulates greater than 100 microns in size.

[0025] In some embodiments, the heating element may be configured to heat an effective amount of the enzyme solution to an effective temperature. In some embodiments, the at least one well may be configured to hold a buffer solution in the range of 1 ml to 100 ml. In some embodiments, the at least one well may be configured to hold a buffer solution in the range of 1 ml to 500 ml. In some embodiments, the at least one well is configured to receive a volume of the enzyme solution.

[0026] In some embodiments, the mortar screen (or screens) may be flat and disposed perpendicular to the sides of the mortar cup (or cups). In some embodiments, the mortar screen (or screens) may be conical. The mortar cup (or cups) may include a mating mortar cup (or cups).

[0027] According to a second aspect, there is provided a method for the automated preparation of a regenerative epithelial suspension, the method may include receiving, by a processor, an initiation signal indicating that a cartridge has been placed on a sensor.

[0028] The cartridge may include (i) a cartridge upper surface having a raised processing area, at least two openings, a mortar cup (or cup) disposed in the raised processing area, a mortar screen (or screen) disposed in the mortar cup (or cup), a tissue sample disposed in the mortar cup (or cup), and a pestle having a pestle head at an end and disposed in the mortar cup (or cup) over the tissue sample; and (ii) a well plate and at least three wells disposed along the well plate, wherein a first well contains a quantity of enzyme, a second well contains a first quantity of buffer, and a third well contains a second quantity of buffer.

[0029] The method may further include starting a housing motor configured to move the pestle shaft (or shafts), and at least one base plate motor configured to rotate the well plate.

[0030] The method may further include performing at least one sequence with the base plate motor and the housing motor, the at least one sequence may include actuating, with the housing motor, a pestle head within the mortar cup (or cups) in the presence of the enzyme solution for an effective time, the actuating may include moving the pestle head up and down along a vertical axis.

[0031] At least one sequence may further include raising the mortar cup (or cup) to an upper position by the housing motor. At least one sequence may further include pressing the pestle head against the tissue sample by the housing motor, the tissue sample may be disposed along the mortar screen (or screen), and the pressing may include pressing the pestle head at least once (e.g., three times), each pressing comprising an effective amount of force.

[0032] At least one sequence may further include rotating the well plate by the base plate motor so that the wells containing the mortar cup (or cup), tissue sample, and pestle head may be in fluid contact with the first amount of buffer. At least one sequence may further include actuating the pestle head in the mortar cup (or cup) in the presence of the buffer solution by the housing motor for an effective time. The actuating may include moving the pestle head up and down along a vertical axis.

[0033] At least one sequence may further include raising the mortar cup (or cup) to an upper position by the housing motor. At least one sequence may further include pressing the pestle head against the tissue sample by the housing motor, and the tissue sample may be disposed along the mortar screen (or screen). The pressing may include pressing the pestle head at least once (e.g., three times). Each pressing may include an effective amount of force.

[0034] At least one sequence may further include rotating the well plate by the base plate motor so that the wells containing the mortar cup (or cup), tissue sample, and pestle head may be in fluid contact with the second amount of buffer. At least one sequence may further include actuating the pestle head within the mortar cup (or cup) in the presence of the second buffer solution by the housing motor. The pestle head may be caused to apply a rotational force and a grinding force to the tissue sample for an effective period of time.

[0035] At least one sequence may further include raising the mortar cup (or cups) to an upper position above the well plate by the housing motor. At least one sequence may further include pressing the pestle head against the tissue sample by the housing motor, and the tissue sample may be disposed along the mortar screen (or screen). The pressing may include pressing the pestle head at least once (e.g., three times). Each pressing may include an effective amount of force.

[0036] The at least one sequence may further include rotating, by a base plate motor, the well plate such that a well containing the second amount of buffer may be disposed directly beneath one of the at least two openings. The at least one sequence may further include receiving, by a sensor, a completion signal that may be configured to indicate that the regenerative epithelial suspension is present in the at least one well.

[0037] In some embodiments, a system for the automated preparation of a regenerative epithelial suspension may comprise a non-transitory tangible computer-readable medium having stored thereon computer-executable instructions that, when executed by a computer processor, cause the system to perform the methods for operating the system as described herein.

[0038] In some examples, a system for automated preparation of a regenerative epithelial suspension may include a base unit including a tissue processing area; a cartridge configured to be received in the tissue processing area, the cartridge including a cover including an opening configured to receive a tissue sample, and a well plate below the cover and configured to rotate relative to the cover; and a tissue disperser configured to mechanically dissociate the tissue when the tissue sample is placed in the cartridge. The system may further include a heating element disposed in the base unit. The heating element may be configured to generate heat sufficient to increase enzymatic efficiency of an enzyme disposed in the well plate. The tissue disperser may include a pestle. The base unit may include a tissue disperser shaft. The tissue disperser shaft may be configured to be attached to the tissue disperser. The system may further include a cup including a screen disposed within the cup. The screen may be laterally oriented within the cup. The opening may include a ridge and be configured to receive the cup. The well plate may include wells configured to align the wells with the openings upon rotation. The well plate may include a plurality of wells and may be configured to rotate to align each well with an opening. The well plate may include a buffer well configured to receive a buffer and an enzyme well configured to receive an enzyme. The well plate may be configured to rotate to position the enzyme well below the opening when an enzyme is used to disperse the tissue. The well plate may be configured to rotate to position the buffer well below the opening when a buffer is applied to the tissue. The system may further include a processor configured to operate the tissue disperser and rotate the well plate.

[0039] In some examples, a system for automated preparation of a regenerative epithelial suspension may include a base unit including a tissue processing area, a cartridge configured to be received in the tissue processing area, the cartridge including a cover with a raised processing opening configured to receive a cup, and a well plate below the cover and configured to rotate relative to the cover, the well plate including wells, and a tissue dissociation device configured to mechanically dissociate tissue when a tissue sample is placed in the cup. The well plate may include at least a first well configured to receive an enzyme solution and a second well configured to receive a buffer solution, and may be configured to rotate to align the first well or the second well with the raised processing opening. The cup may be configured to be lowered and raised into the well of the well plate.

[0040] 19. The system of any one of claims 16 to 18, wherein the base unit comprises a heating element disposed within the housing.

[0041] 20. The system of any one of claims 16 to 19, wherein the cartridge comprises a cup and a screen disposed within the cup.

[0042] In certain examples, a cartridge for preparing a regenerative epithelial suspension may include a cover with a raised processing opening, a cup configured to be received within the raised processing opening and including a screen, and a well plate disposed below the cover, including wells, configured to align the wells with the raised processing opening upon rotation relative to the cover. The screen may be laterally disposed within the cup. The screen may be configured to separate particulates greater than 100 microns in size. The screen may be configured to separate particulates greater than 50 microns in size. The screen may be configured to separate particulates greater than 10 microns in size. The cartridge may further include a docking spindle configured to connect to a base unit. The docking spindle may be configured to rotate the well plate. The wells may include raised processing elements configured to enhance dispersion of tissue samples. The raised processing elements may be configured to enhance dispersion of tissue samples subjected to normal, rotational, or grinding forces. The cup may be configured to be positioned within the wells by raising or lowering within the raised processing opening. The cover may include multiple openings. The cover may include a buffer opening. The well plate may include a plurality of openings. The well plate may include a buffer well and an enzyme well. The well plate may be configured to rotate to position the buffer well directly below the buffer opening. The well plate may be configured to rotate to position the enzyme well directly below the raised treatment opening. The cup may be configured to receive a tissue dispersal device.

[0043] In some examples, a cartridge for preparing a regenerative epithelial suspension may include a cover having an opening configured to receive a tissue sample, and a well plate disposed below the cover, including wells, configured to align the wells with the openings upon rotation relative to the cover. The cartridge may be configured to lower the tissue sample into the wells aligned with the openings. The well plate may include buffer wells and enzyme wells, configured to align the buffer wells or enzyme wells with the openings upon rotation. The cartridge may further include a cup receivable within the opening in the cover and movable between a raised position above the wells and a lowered position within the wells.

[0044] In certain examples, a method for automated preparation of a regenerative epithelial suspension may include receiving, by a processor, a start signal indicating that a cartridge including a cover, a cup containing a tissue sample, a well plate having a first well directly below the cover and containing a volume of enzyme solution, and a second well containing a volume of buffer solution has been placed on a sensor. The method may include executing a sequence including: operating a tissue disperser on the tissue sample in the presence of the enzyme solution with the cup positioned in the first well; raising the cup to an upper position; rotating the well plate to position the second well directly below the cup; lowering the cup to a lower position within the second well; and operating the tissue disperser on the tissue sample in the presence of the buffer solution with the cup positioned in the second well. The tissue disperser may apply a rotational force to the tissue sample. The tissue disperser may apply a grinding force to the tissue sample. The tissue disperser may include a pestle. The tissue disperser may include a raised surface element. The tissue disperser may include multiple protrusions. The tissue sample may be disposed along a screen disposed within the cup. The screen may be disposed laterally within the cup. The screen may be configured to separate particulates greater than 100 microns in size. The screen may be configured to separate particulates greater than 50 microns in size. The screen may be configured to separate particulates greater than 10 microns in size.

[0045] In some examples, a method for automated preparation of a regenerative epithelial suspension (as described in the preceding paragraph) may include starting a housing motor configured to operate a tissue dispersion device. The housing motor may be configured to raise and lower a cup. The method may further include pressing the tissue dispersion device against the tissue sample multiple times when the cup is disposed in the first well and the second well. The method may further include operating the tissue dispersion device against the tissue sample when the cup is in the upper position. The method may further include starting a base plate motor configured to rotate the well plate. The method may further include bringing the cup into fluid contact with a second amount of buffer solution upon rotation of the well plate. The second amount of buffer solution may be contained in a third well of the well plate. The method may further include operating the tissue dispersion device against the tissue sample in the presence of the second amount of buffer solution with the cup disposed in the third well, the third well comprising one or more raised processing elements. The method may further include receiving a completion signal indicating the presence of a regenerative epithelial suspension. The tissue disperser may be adapted to operate on the tissue sample in the presence of an enzyme solution with the cup disposed in the first well for a time effective to at least partially dissociate the tissue sample. The tissue disperser may be adapted to operate on the tissue sample in the presence of an enzyme solution with the cup disposed in the first well for a time effective to at least partially dissociate the tissue sample. The tissue disperser may be adapted to operate on the tissue sample in the presence of a buffer solution with the cup disposed in the second well with an amount of force effective to at least partially dissociate the tissue sample. In some embodiments, a non-transitory tangible computer-readable medium is provided having stored thereon computer-executable instructions that, when executed by a computer processor, cause the method to be performed as described in this section or elsewhere herein.

[0046] In certain examples, a method for treating a tissue site may include collecting a tissue sample containing keratinocytes and placing it in a cartridge including a well plate; placing the cartridge in a base unit including a tissue dispersion device; and actuating the base unit to rotate the well plate and operate the tissue dispersion device, the well plate rotating to align one or more wells in the well plate with the tissue sample and the tissue dispersion device, and operating the tissue dispersion device to form a regenerative epithelial suspension by dissociating the tissue sample; and delivering the regenerative epithelial suspension to the tissue site to enhance healing of the tissue site. The tissue sample may include a skin sample. The regenerative epithelial suspension may include a mixed population of viable cells. The regenerative epithelial suspension may include fibroblasts. The regenerative epithelial suspension may include melanocytes. The tissue site may be a burn tissue site. The tissue site may be a full-thickness skin defect. The cartridge may include a cover including a raised treatment opening. The method may further include placing the tissue sample in a cup disposed within the raised treatment opening. Activation of the base unit may cause the cup to lower into the well of the well plate. In certain examples, the tissue dissociation device may be a pestle. The well plate may include an enzyme well. The method may include disposing an enzyme configured to dissociate tissue in the enzyme well. The well plate may be positioned such that the enzyme well is positioned directly beneath the tissue sample. Activation of the base unit may cause the tissue aggregator to act on the tissue sample in the presence of the enzyme in the enzyme well. The well plate may include a buffer well. The method may include disposing a buffer in the buffer well. Activation of the base unit may cause rotation of the well plate such that the buffer well is positioned directly beneath the tissue sample. Activation of the base unit may cause the tissue aggregator to act on the tissue sample in the presence of the buffer in the buffer well.The well may include one or more raised treatment elements. [Brief explanation of the drawings]

[0047] [Figure 1] FIG. 1 is a front perspective view of a system for automated preparation of regenerative epithelial suspensions according to one or more embodiments of the present disclosure. [Figure 2] FIG. 1 illustrates a front perspective view of a base unit according to one or more embodiments of the present disclosure. [Figure 3A] FIG. 1 is an exploded perspective view of a cartridge assembly according to one or more embodiments of the present disclosure. [Figure 3B] FIG. 10 is a top perspective view of a cartridge top cover according to one or more embodiments of the present disclosure. [Figure 3C] FIG. 13 is a bottom perspective view of a cartridge top cover according to one or more embodiments of the present disclosure. [Figure 3D] FIG. 1 is a top perspective view of a well plate according to one or more embodiments of the present disclosure. [Figure 3E] FIG. 10 is a top perspective view of an alternative embodiment of a well plate in accordance with one or more embodiments of the present disclosure. [Figure 3F] FIG. 1 is a bottom perspective view of a well plate according to one or more embodiments of the present disclosure. [Figure 3G] FIG. 10 is a top perspective view of a cartridge bottom cover according to one or more embodiments of the present disclosure. [Figure 3H] FIG. 10 is a bottom perspective view of a cartridge bottom cover according to one or more embodiments of the present disclosure. [Figure 4] FIG. 1 is an exploded perspective view of a pestle assembly according to one or more embodiments of the present disclosure. [Figure 5A] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5B] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5C]FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5D] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5E] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5F] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5G] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5H] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5I] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 5J] FIG. 1 is a diagram generally illustrating one non-limiting embodiment of a pestle head according to the present disclosure. [Figure 6] 1 is a flowchart illustrating an automated sequence and associated structural components according to one or more embodiments of the present disclosure. [Figure 7A] 1 is a diagram generally disclosing one non-limiting embodiment of a mortar cup (or cup) according to the present disclosure. [Figure 7B] 1 is a diagram generally disclosing one non-limiting embodiment of a mortar cup (or cup) according to the present disclosure. [Figure 7C] 1 is a diagram generally disclosing one non-limiting embodiment of a mortar cup (or cup) according to the present disclosure. [Figure 8A] 1 is a diagram generally disclosing a combination of a mating pestle, a mating mortar cup (or cup), and a mating well C. [Figure 8B] 10 is a diagram generally disclosing an alternative combination of a mating pestle, a mating mortar cup (or cup), and a mating well C. [Figure 9A] FIG. 10 is a top view of a mating well C containing a first embodiment of a raised treatment element. [Figure 9B]FIG. 10 is a top view of a mating well C containing a second embodiment of a raised treatment element. [Figure 9C] FIG. 10 is a side perspective view of the mating well C. [Figure 10A] FIG. 1 is a perspective view of one four-piece embodiment of packaging for a set of tools including two clamshell tool set pairs and a bottom tray configured to receive the set of tools. [Figure 10B] FIG. 10 is a top view of the upper clamshell component of the first clamshell toolset pair. [Figure 10C] FIG. 10 is a bottom view of the upper clamshell component of the first clamshell tool set pair. [Figure 10D] FIG. 10 is a top view of the lower clamshell component of the first clamshell toolset pair. [Figure 10E] FIG. 10 is a bottom view of the lower clamshell component of the first clamshell tool set pair. [Figure 10F] FIG. 10 is a top view of the upper clamshell component of the second clamshell tool set pair. [Figure 10G] FIG. 10 is a bottom view of the upper clamshell component of the second clamshell tool set pair. [Figure 10H] FIG. 10 is a top view of the lower clamshell component of the second clamshell tool set pair. [Figure 10I] FIG. 10 is a bottom view of the lower clamshell component of the second clamshell tool set pair. [Figure 10J] FIG. 10 is a top view of the lower clamshell component of the first clamshell toolset pair shown containing a tool. [Figure 10K] FIG. 10 is a top view of the lower clamshell component of the second clamshell toolset pair shown containing a tool. [Figure 11] A top, front, right-side perspective view of the base unit. [Figure 12] FIG. 2 is a front view of the base unit. [Figure 13] FIG. 2 is a rear view of the base unit. [Figure 14] FIG. 2 is a left side view of the base unit. [Figure 15] FIG. 2 is a right side view of the base unit. [Figure 16] FIG. 2 is a top view of the base unit. [Figure 17] FIG. 2 is a bottom view of the base unit. [Figure 18] Top, front, right-side perspective view of the cartridge. [Figure 19] FIG. [Figure 20] FIG. [Figure 21] FIG. [Figure 22] FIG. [Figure 23] FIG. [Figure 24] FIG. DETAILED DESCRIPTION OF THE INVENTION

[0048] Having generally described various aspects of the present disclosure, reference will now be made in detail to the subject matter illustrated in the drawings. While the present disclosure will be described with reference to these drawings, it is not intended to be limited to one or more embodiments disclosed herein. Rather, it is intended to cover all alternatives, modifications, and equivalents included within the spirit and scope of the present disclosure as defined by the appended claims. For purposes of overview, certain aspects, advantages, and novel features have been described. However, it is understood that not all such advantages may be achieved in accordance with any particular embodiment. Thus, the disclosed subject matter may be embodied or practiced to achieve or optimize a certain advantage or group of advantages without achieving every advantage that may be taught or suggested.

[0049] The devices, apparatus, compositions, suspensions, methods, and systems described herein may be adapted for use in treating a tissue site, which may be a wound site, a burn site, a full-thickness skin defect, a vitiligo lesion, a site undergoing and / or suitable for repigmentation, a user- or surgeon-inflicted wound, trauma, and / or any suitable injury or defect.

[0050] In some embodiments, one or more systems for automated preparation of regenerative epithelial suspensions include a base unit, a cartridge, and a tissue disperser. Those skilled in the art will appreciate that tissue separation and / or dispersion can be achieved using a tissue disperser. The tissue disperser can take many forms, such as a pestle, a grinder, a blade, various cutting and blunt instruments, a cutting screen, or any suitable tool. While reference is made throughout this specification to a pestle, any suitable tissue disperser may be used. In some embodiments, the system may further include a set of associated tools.

[0051] In some embodiments, the base unit may include a housing, a front cover, and a tissue processing area. In some embodiments, the housing may include one or more side panels, a housing top, a display having one or more buttons, a touchscreen, or both one or more buttons and a touchscreen, a base front plate, and two or more base foot pads. The base unit may further include a pestle shaft (or shaft), a power supply, one or more circuit boards having one or more memory units, and one or more motors. In some embodiments, the tissue processing area may include one or more heating elements, a docking spindle, one or more lighting elements, one or more side cartridge alignment protrusions, one or more rear cartridge alignment protrusions, and one or more control feedback sensors.

[0052] The cartridge assembly (or simply "cartridge" herein) may include a cartridge top cover, a raised processing area, one or more upper processing shelves, "A," "B," "C," and "D" openings in the cartridge top cover, one or more cartridge tabs, a mortar top cap, a mortar polymer ring, a mortar cup (or cup), a mortar screen (or screen), a well plate, "A," "B," and "C" wells in the well plate, one or more raised processing elements in the "C" well, a well heating cup disposed directly below the "A" well in the well plate, a drive sleeve, a cartridge bottom cover, a cartridge tip, a bottom cover opening in the cartridge bottom cover, and a docking spindle opening at the bottom cover opening in the cartridge bottom cover. Those skilled in the art will understand that the term "mortar cup" is synonymous with the word "cup" and does not indicate that a pestle must be used with the cup. As would be otherwise understood by one of ordinary skill in the art, any suitable container or any suitable receiving receptacle may be substituted for the mortar cup or cup in this section and throughout this specification. Additionally, one of ordinary skill in the art will appreciate that the term "mortar screen" is synonymous with the term "screen" and does not indicate that a pestle must be used in conjunction with the screen. As would be otherwise understood by one of ordinary skill in the art, any suitable screen may be substituted for the mortar screen in this section and throughout this specification.

[0053] The pestle assembly (or simply "pestle" herein) may include a pestle top cap, a pestle spring, one or more pestle retention wires, a pestle spring cap, a pestle bottom cap, a pestle body, and a pestle head. In some embodiments, the pestle may further include a pestle shaft (or shaft).

[0054] As may be more fully understood herein, the pestle head of the present disclosure may comprise at least one outer surface and a distal surface (or "terminal end" as used herein). In some embodiments, the terminal end is flat. In some embodiments, the terminal end is dome-shaped.

[0055] In some embodiments, the distal end includes a mating end from which extends a column distally, the column having a smaller circumference than the outer surface. In this regard, in some embodiments, the mortar cup (or cup) may include a sidewall, a distal opening, and a screen disposed across the distal opening. In some embodiments, the distal opening and screen of the mortar cup (or cup) may be configured perpendicular to the sidewall, corresponding to the flat end embodiment of the pestle head. In some embodiments, the distal opening and screen of the mortar cup (or cup) may be curved, corresponding to the domed end embodiment of the pestle head. In some embodiments, the distal opening and screen of the mortar cup (or cup) may be configured with a mating opening across which the screen is disposed, corresponding to the mating end embodiment of the pestle head. In this regard, in some embodiments, the raised portion of one or more wells (such as, but not limited to, well C) may be generally planar, curved, or mating so as to align with the planar, domed, or mating pestle head and associated planar, curved, or mating distal end and configuration of the mortar cup (or cups).

[0056] Tools that accompany the cartridge and pestle include, but are not limited to, one or more vials of buffer solution, one or more vials of enzyme solution, one or more vials of sterile water, one or more syringes, one or more needles, one or more scalpels, one or more labels, and one or more spray nozzles.

[0057] In some embodiments, an automated method for preparing a regenerative epithelial suspension may include (i) starting location identification, (ii) user settings, and (iii) one or more tissue processing sequences. One or more methods disclosed herein may further include post-processing by a user.

[0058] In embodiments, a system for automated preparation of regenerative epithelial suspension may first verify the following starting positions: the mortar cup (or cups) is on one or more upper processing shelves in the cartridge, the well plate is in the "A" position, and the pestle shaft (or shafts) is raised.

[0059] In some embodiments, user setup may include the steps of: (a) powering on the system; (b) inserting a predetermined amount of buffer into well B (buffer well) and well C through openings "B" and "C" respectively in the cartridge top cover; (c) inserting a predetermined amount of enzyme solution into well A (enzyme well) through opening "A" in the cartridge top cover; (d) obtaining a tissue sample; (e) placing the tissue sample in a mortar cup (or cup) in the raised processing area of ​​the cartridge top cover; (f) placing a pestle on the tissue sample in the mortar cup (or cup) in the raised processing area of ​​the cartridge top cover; (g) lifting the front cover; (h) placing the cartridge in the tissue processing area and aligning the cartridge tip with one or more control feedback sensors and the heating element with the well heating cup; (i) closing the front cover; and (j) pressing one or more buttons on the display to start processing.

[0060] In embodiments, the system may execute one or more tissue processing sequences, including: (a) a pestle shaft (or shaft) may be lowered and inserted into the pestle cap through an opening in the top surface of the pestle cap; (b) a pestle shaft (or shaft) may be rotated to engage with the pestle body; (c) one or more raised elements disposed along the outer surface of the pestle body may be engaged with one or more raised elements disposed along the inner surface of the pestle head; and (d) a pushing, rotating, or both pushing and rotating of the pestle body by the pestle shaft (or shaft) may cause one or more raised side elements disposed on the exterior of the pestle head to contact the inner surface of the mortar cup (or cup), causing the mortar cup (or cup) to slide off one or more upper processing shelves and lower into well A (enzyme well). When the mortar cup (or cup) is in Well A (enzyme well), the enzyme solution may pass through the mortar screen (or screen) to contact the tissue sample. (e) A heating element may heat the well heating cup, which may heat the enzyme solution in Well A (enzyme well). (f) The pestle shaft (or shaft) may move the pestle vertically up and down in a predetermined sequence. (g) After a predetermined time, the pestle shaft (or shaft) may rotate the pestle, causing the pestle to rotate the mortar cup (or cup) in the opposite direction and raise the mortar cup (or cup) to the "upper" position. When the mortar cup (or cup) is in the "upper" position, the edge of the mortar cup (or cup) resides on one or more upper processing shelves. (h) After a predetermined time, the pestle shaft (or shaft) may press the pestle down three times, causing the pestle to press the tissue sample against the mortar screen (or screen). Such pressing may be performed with a predetermined pound of force sufficient to squeeze the enzyme solution out of the tissue sample.(i) The well plate may rotate to align well B (buffer well) directly below the raised processing area. (j) The pestle shaft (or shaft) may rotate the pestle, causing the pestle head to engage the mortar cup (or cup) and slide the mortar cup (or cup) down from one or more upper processing shelves into well B (buffer well). When the mortar cup (or cup) is in well B (buffer well), the buffer solution may pass through the mortar screen (or screen) to contact the tissue sample. (k) The pestle shaft (or shaft) may move the pestle vertically up and down in a predetermined sequence. (l) After a predetermined time, the pestle shaft (or shaft) may rotate the pestle, causing the pestle to rotate the mortar cup (or cup) in the opposite direction and raise the mortar cup (or cup) to the "upper" position. When the mortar cup (or cup) is in the "up" position, the edge of the mortar cup (or cup) rests on one or more upper processing shelves. (m) After a predetermined time, the pestle shaft (or shaft) may press the pestle three times downward, causing the pestle to press the tissue sample against the mortar screen (or screen). Such pressing may be performed with a predetermined pound of force sufficient to squeeze the buffer solution out of the tissue sample. (n) The well plate may rotate to align well C directly below the raised processing area. (o) The pestle shaft (or shaft) may rotate the pestle body, causing the pestle head to engage the mortar cup (or cup), sliding the mortar cup (or cup) down from one or more upper processing shelves into well C. When the mortar cup (or cup) is in well C, the buffer solution may pass through the mortar screen (or screen) to contact the tissue sample. (p) The pestle shaft (or shafts) may be rotated in a predetermined sequence such that the pestle head exerts a rotational and grinding force on the tissue sample in the presence of a buffer solution.The mortar screen (or screen) and raised processing element may have surfaces on which the tissue sample can be subjected to rotational and grinding forces, providing an associated counter pressure. (q) After a predetermined time, the pestle shaft (or shaft) may rotate the pestle body and pestle head, causing the pestle to rotate the mortar cup (or cup) in the opposite direction and raise the mortar cup (or cup) to the "up" position. When the mortar cup (or cup) is in the "up" position, the edge of the mortar cup (or cup) rests on one or more upper processing shelves. (r) After a predetermined time, the pestle shaft (or shaft) may press the pestle three times downward, causing the pestle to press the tissue sample against the mortar screen (or screen). These pressings may be performed with a predetermined pound of force sufficient to squeeze the buffer solution out of the tissue sample. (s) The well plate may rotate to align well C directly under the "D" opening in the cartridge top cover. (t) The sensors may send signals to one or more memory units to record processing events.

[0061] In embodiments in which the pestle shaft (or shaft) comprises part of a pestle, the above sequence may alternatively include: (a) an ejection element may descend from the top of the housing to removably engage with the pestle shaft (or shaft); (b) the pestle shaft (or shaft) may be adapted to rotate to engage with the pestle body; (c) one or more raised elements disposed along the outer surface of the pestle body may be adapted to engage with one or more raised elements disposed along the inner surface of the pestle head; and (d) pressing, rotating, or both pressing and rotating the pestle body by the pestle shaft (or shaft) may cause one or more raised side elements disposed on the exterior of the pestle head to contact the inner surface of the mortar cup (or cup), causing the mortar cup (or cup) to slide down from one or more upper processing shelves into well A. When the mortar cup (or cup) is in well A, the enzyme solution may pass through the mortar screen (or screen) to contact the tissue sample. (e) A heating element may heat the well heating cup, which in turn heats the enzyme solution in well A. (f) The pestle shaft (or shaft) may move the pestle vertically up and down in a predetermined sequence. (g) After a predetermined time, the pestle shaft (or shaft) may rotate the pestle, causing the pestle to rotate the mortar cup (or cup) in the opposite direction and raise the mortar cup (or cup) to the "upper" position. When the mortar cup (or cup) is in the "upper" position, the edge of the mortar cup (or cup) resides on one or more upper processing shelves. (h) After a predetermined time, the pestle shaft (or shaft) may press the pestle down three times, causing the pestle to press the tissue sample against the mortar screen (or screen). Such pressing may be performed with a predetermined poundage of force sufficient to squeeze the enzyme solution out of the tissue sample. (i) The well plate may be rotated to align well B (the buffer well) directly beneath the raised treatment area.(j) The pestle shaft (or shaft) may rotate the pestle, causing the pestle head to engage with the mortar cup (or cup), sliding the mortar cup (or cup) down from one or more upper processing shelves into well B. When the mortar cup (or cup) is in well B, the buffer solution may pass through the mortar screen (or screen) to contact the tissue sample. (k) The pestle shaft (or shaft) may move the pestle vertically up and down in a predetermined sequence. (l) After a predetermined time, the pestle shaft (or shaft) may rotate the pestle, causing the pestle to rotate the mortar cup (or cup) in the opposite direction, raising the mortar cup (or cup) to an "upper" position. When the mortar cup (or cup) is in the "upper" position, the edge of the mortar cup (or cup) resides on one or more upper processing shelves. (m) After a predetermined time, the pestle shaft (or shaft) may press the pestle three times downward, causing the pestle to press the tissue sample against the mortar screen (or screen). Such pressing may be performed with a predetermined pound of force sufficient to squeeze the buffer solution out of the tissue sample. (n) The well plate may rotate to align well C directly below the raised processing area. (o) The pestle shaft (or shaft) may rotate the pestle body, causing the pestle head to engage with the mortar cup (or cups) and slide the mortar cup (or cups) down from one or more upper processing shelves into well C. When the mortar cup (or cups) is in well C, the buffer solution may pass through the mortar screen (or screen) to contact the tissue sample. (p) The pestle shaft (or shaft) may rotate in a predetermined sequence, causing the pestle head to apply rotational and grinding forces to the tissue sample in the presence of the buffer solution.The mortar screen (or screen) and raised processing element may have surfaces on which the tissue sample may be subjected to rotational and grinding forces, exerting associated opposing pressures. (q) After a predetermined time, the pestle shaft (or shaft) may rotate the pestle body and pestle head, causing the pestle to rotate the mortar cup (or cup) in the opposite direction and raise the mortar cup (or cup) to an "upper" position. When the mortar cup (or cup) is in the "upper" position, the edge of the mortar cup (or cup) rests on one or more upper processing shelves. (r) After a predetermined time, the pestle shaft (or shaft) may press the pestle three times downward, causing the pestle to press the tissue sample against the mortar screen (or screen). These pressings may be performed with a predetermined pound of force sufficient to squeeze buffer solution out of the tissue sample. (s) The well plate may rotate to align well C directly under the "D" opening in the cartridge top cover. (t) The sensors may send signals to one or more memory units to record processing events.

[0062] Post-treatment by the user may include the steps of: (a) lifting the front cover; (b) inserting one or more syringes into well C through opening "D" in the cartridge top cover; (c) drawing the regenerative epithelial suspension up into the syringe; and (d) treating one or more treatment areas of the patient with the regenerative epithelial suspension. In some embodiments, treating the patient with the regenerative epithelial suspension may include replacing the syringe needle with a spray nozzle and then spraying the regenerative epithelial suspension onto the treatment site.

[0063] After processing, the system may raise the pestle shaft (or shaft) through an opening in the pestle top cap, allowing the user to remove the used cartridge and pestle. In embodiments where the pestle additionally comprises a pestle shaft (or shaft), an ejection element may release the pestle shaft (or shaft), allowing the user to remove the used cartridge and pestle.

[0064] In some embodiments, a system for automated preparation of regenerative epithelial suspension, when executed by a computer processor, includes: (a) receiving, by the processor, a start signal indicating that a cartridge has been placed on a sensor, the cartridge including: (i) a cartridge top surface having a raised processing area, at least two openings, a mortar cup (or cup) disposed in the raised processing area, a mortar screen (or screen) disposed in the mortar cup (or cup), a tissue sample disposed in the mortar cup (or cup), and a pestle having a pestle head at an end and disposed in the mortar cup (or cup) over the tissue sample; (ii) a well plate and at least three wells disposed along the well plate, wherein a first well contains an amount of enzyme, a second well contains a first amount of buffer, and a third well contains a second amount of buffer; and (b) starting a housing motor configured to operate the pestle shaft (or shaft). (c) starting at least one base plate motor configured to rotate the well plate; and (d) using the base plate motor and the housing motor, (i) actuating a pestle head in the mortar cup (or cup) in the presence of the enzyme solution by the housing motor for an effective time, said actuating comprising moving the pestle head up and down along a vertical axis; (ii) raising the mortar cup (or cup) to an upper position by the housing motor, said raising comprising actuating a pestle shaft (or shaft) which actuates the pestle such that the pestle is releasably engaged with the mortar cup (or cup) by contact between one or more raised side elements disposed on the pestle head and an inner surface of the mortar cup (or cup), said contact being sufficient to allow lifting, rotation or manipulation of the mortar cup (or cup) by the pestle; and (iii) actuating a mortar screen (or(iv) rotating the well plate by the base plate motor so that the wells containing the mortar cup (or cup), the tissue sample, and the pestle head are in fluid contact with a first amount of buffer; (v) actuating the pestle head in the mortar cup (or cup) in the presence of the buffer solution for an effective time by the housing motor, the actuating comprising moving the pestle head up and down along a vertical axis; (vi) raising the mortar cup (or cup) to an upper position by the housing motor; (vii) actuating the pestle head by the housing motor against the tissue sample disposed along the mortar screen (or screen), the actuating comprising pressing the pestle head three times, each pressing comprising an effective amount of force; (viii) rotating the well plate by the base plate motor so that the wells containing the mortar cup (or cups), tissue samples, and pestle heads are in fluid contact with the second amount of buffer; (ix) actuating the pestle head in the mortar cup (or cups) in the presence of the second buffer solution by the housing motor so that the pestle head exerts rotational and grinding forces on the tissue sample for an effective time; (x) raising the mortar cup (or cups) by the housing motor to an upper position above the well plate; (xi) pressing the pestle head against the tissue sample disposed along the mortar screen (or screen) by the housing motor, said pressing comprising pressing the pestle head three times, each pressing comprising an effective amount of force; (xii) rotating the well plate by the base plate motor so that the wells containing the second amount of buffer are disposed directly below one of the at least two openings; (xiii) detecting by the sensor;and receiving a completion signal configured to indicate that the regenerative epithelial suspension is present in at least one well.

[0065] Those skilled in the art will appreciate that the start signal may be sent when the sensor detects the presence or absence of a cartridge, which may or may not initiate the sequence. However, in some examples, a user may initiate the sequence with a start signal, such as by using a button, tap, switch, or any suitable mechanism. In some examples, this signal may only be effective if a start signal is sent from the sensor.

[0066] In some embodiments, the predetermined pounds of force sufficient to express the solution from the tissue sample may include 1-10 pounds of force, 2-5 pounds of force, 10-20 pounds of force, 20 pounds of force, or greater than 20 pounds of force.

[0067] One or more embodiments of the present disclosure may be implemented as a program product for use with a computer system. The program of the program product defines the functions of the embodiments (including the methods described herein) and may be contained in a variety of computer-readable storage media. Exemplary computer-readable storage media include, but are not limited to, (i) a writable storage medium on which changeable information is stored (e.g., a portable drive such as a hard disk, USB stick, or floppy disk) or (ii) a non-writable storage medium on which information is permanently stored (e.g., a read-only memory device within a computer, such as a CD-ROM disk readable by a CD-ROM drive). Such computer-readable storage media are embodiments of the present invention when they contain computer-readable instructions that direct the functions of the present invention. Other media include communication media through which information is delivered to a computer, such as computer or telephone networks, including wireless communication networks. The latter embodiment specifically includes the transmission of information over the Internet and other networks. Such communication media are embodiments of the present invention when they contain computer-readable instructions that direct the functions of the present invention. Computer-readable storage media and communication media may be broadly referred to herein as computer-readable media. In a preferred embodiment, the computer-readable storage medium of the present invention may comprise one or more printed wiring board assemblies communicatively coupled to one or more memory units housed within a housing of the present invention.

[0068] Generally, the routines executed to implement embodiments of the present invention may be part of an operating system or a specific application, component, program, module, object, or instruction sequence. A computer program of the present invention is typically made up of a number of instructions that are translated by a native computer into a machine-readable format and, therefore, executable instructions. The program is also made up of variables and data structures that either reside locally to the program or are found in memory or storage devices. Various programs described below may also be identified based on the application for which they are implemented in a specific embodiment of the present invention. It should be understood, however, that any specific program names below are used for convenience only, and the present invention should not be limited to use with any specific application identified and / or implied by such names.

[0069] For simplicity and clarity of illustration, the drawings show general configurations, and descriptions and details of well-known features and techniques may be omitted so as not to unnecessarily obscure the present invention. Furthermore, the drawings of elements in the drawings are not necessarily to scale. For example, the dimensions of some elements in the drawings may be exaggerated relative to other elements to help improve understanding of embodiments of the present invention. Furthermore, the same reference numerals in different drawings refer to the same elements.

[0070] In this specification and claims, when the terms "first," "second," "third," "fourth," etc. appear, they are used to distinguish between similar elements and do not necessarily represent a particular sequence or chronological order. It is understood that the interchange of terms used in this manner, under appropriate circumstances, may, for example, enable the embodiments described herein to operate in sequences other than those shown or described herein. Furthermore, the terms "include" and "have," and any variations of each, are intended to cover non-exclusive inclusions, and a process, method, system, article, device, or apparatus comprising a list of elements is not necessarily limited to those elements but may include other elements not expressly listed or inherent to such process, method, system, article, device, or apparatus.

[0071] The term "couple" (e.g., "coupled," "couples," "coupling," etc.) should be understood broadly and refers to the electrical, mechanical, or other connection of two or more elements or signals. Two or more electrical elements may be electrically coupled but not mechanically or otherwise coupled; two or more mechanical elements may be mechanically coupled but not electrically or otherwise coupled; two or more electrical elements may be mechanically coupled but not electrically or otherwise coupled. The coupling (whether mechanical, electrical, or otherwise) may be of any duration, e.g., permanent, semi-permanent, or momentary.

[0072] As used herein, the term "digital" refers to any operation, version, configuration, expression, or other element that resides primarily or exclusively in a computer program or electronic medium.

[0073] As used herein, "effective" (including, but not limited to, an effective time or an effective amount of one or more solutions) means an amount sufficient to accomplish the intended task. By way of non-limiting example, an effective amount of enzyme solution may comprise 1-100 ml of enzyme solution, and an effective amount of buffer solution may comprise 1-200 ml of buffer solution. Similarly, in some embodiments, and also by way of non-limiting example, an effective time for contacting a tissue sample with the enzyme solution may comprise 5-30 minutes, in some embodiments, 5-45 minutes, and in some embodiments, 5-60 minutes or more. In some embodiments, an effective time for heating the enzyme solution may be 1-15 minutes, in some embodiments, 1-30 minutes, or longer than 30 minutes. In some embodiments, the enzyme solution may comprise a premix solution comprising at least a certain amount of enzyme in a buffer solution along with one or more additional ingredients.

[0074] In some embodiments, the enzymes in the enzyme solution include, but are not limited to, one or more of trypsin, dispase, collagenase, trypsin-EDTA, thermolysin, pronase, hyaluronidase, elastase, papain, and pancreatin. In some variations, the one or more enzymes, such as trypsin, dispase, collagenase, trypsin-EDTA, thermolysin, pronase, hyaluronidase, elastase, papain, and pancreatin, may be conventionally sourced, for example, by fermentation, or may be recombinant or animal-derived. In some variations, the enzyme solution may be formed by mixing lyophilized enzymes with an appropriate volume of fluid (e.g., water). In one or more embodiments, the enzyme may include recombinant trypsin, dispase, collagenase, trypsin-EDTA, thermolysin, pronase, hyaluronidase, elastase, papain, or pancreatin enzymes, or two or more such recombinant enzymes. When trypsin is used as the enzyme, the enzyme solution is preferably calcium and magnesium free.

[0075] Figure 1 generally discloses one embodiment of a system 101 for the automated preparation of a regenerative epithelial suspension. The various elements of Figure 1 are described in more detail below.

[0076] FIG. 2 illustrates one embodiment of a base unit according to one embodiment of the present disclosure, and more specifically illustrates a base unit 200, a housing 201, a tissue processing area 202, a housing base 203, a front cover 204, a heating element 205, a docking spindle 206, a display 207, a first menu button 208, a second menu button 209, a side cartridge alignment protrusion 210, a rear cartridge alignment protrusion 211, a base front plate 212, a base pad 213, a housing top surface 214, and a control feedback sensor 215.

[0077] As seen in FIG. 2 , in at least one embodiment, it is contemplated that base unit 200 and its housing 201 may generally comprise a countertop-sized unit extending from housing top surface 214 to its base and ultimately to base pad 213. In some embodiments, the front end of housing 201, extending from display 207 to the base front plate, may be generally planar, as seen in FIG. 2 . Additionally, in some embodiments, two, three, four, five, or more base pads 213 are contemplated. Base unit 200 may further comprise one or more computer processing elements, including, but not limited to, one or more printed wiring board assemblies, one or more hard drives, one or more wiring boards, one or more motherboards, one or more central processing units, one or more computer memory elements, one or more random access memories, and one or more processors (not shown). Furthermore, in addition to or instead of the shape of base unit 200 generally shown in FIGS. 1 and 2 , base unit 200 may include shapes such as a cylinder, cube, sphere, cone, rectangular parallelepiped, hexagonal prism, or any other shape that enables the practice of the present disclosure. As one skilled in the art would appreciate, in such embodiments, certain relevant elements may have different shapes. As a non-limiting example, if base unit 200 is shaped as a cylinder, front cover 204 and display 207 may be curved accordingly. Similarly, it is contemplated that such relevant configuration or design variations may be utilized for the practice of the present invention across various embodiments. In some embodiments, base unit 200 itself may be comprised of various subassemblies that can be used in conjunction with each other to produce one or more regenerative epithelial suspensions.

[0078] 2, the heating element 205 is generally depicted as a metal plate configured to conduct heat through a well heating cup 318, as discussed in more detail below, although in some embodiments other heating elements are contemplated, including, but not limited to, chemical heating elements, such as a blister pack that heats upon activation, radiation heating elements, or other heating elements known in the art.

[0079] In some embodiments, the front cover 204 may include a flat cover. In some embodiments, the front cover 204 may include a rounded cover. In some embodiments, the front cover 204 may be generally shaped as three sides of a square. In some embodiments, the front cover 204 may taper downward such that the top may be narrower than the bottom. Conversely, in some embodiments, the top of the front cover 204 may be wider than the bottom of the front cover 204. In some embodiments, the front cover 204 may be translucent, transparent, or "see-through." In some embodiments, the front cover 204 may be partially translucent and partially opaque. In some embodiments, the front cover 204 may be opaque. In some embodiments, the front cover 204 may be omitted.

[0080] As can be seen by comparing generally the cartridge top cover 301 described below, and more specifically the cartridge tab 308, a user can appreciate that in some embodiments, the cartridge alignment protrusion 210 and rear cartridge alignment protrusion 211 can be configured to hold the cartridge 300 in place during one or more processing sequences. In embodiments in which the cartridge top cover 301 has different heights, the cartridge alignment protrusion 210 and rear cartridge alignment protrusion 211 can be positioned low, high, or one high and the other low in the tissue processing area 202. In one or more alternative embodiments, a tab or protrusion can extend from the front cover 204 to releasably engage the cartridge alignment protrusion 210.

[0081] In some embodiments, the docking spindle 206 may be configured to align with the drive sleeve 319. However, in some embodiments, the docking spindle 206 may be any shape that may be advantageous for engagement with the drive sleeve 319 or equivalent element that may be configured to rotate the well plate 313. By way of non-limiting example, the docking spindle 206 or equivalent element may be adapted to engage with the drive sleeve 319 or equivalent element, and may be adapted to reversibly engage via a magnetic connection engagement, a Luer lock, a snap fit engagement, a bayonet-type engagement, an air hose quick connect engagement, a press fit engagement, a "Mix2Vial"® engagement, and any other form of temporary, reversible, or in some embodiments, permanent engagement method known in the relevant art.

[0082] In some embodiments, the control feedback sensor 215 may include a temperature sensor, an electromechanical sensor, a pressure sensor, a vibration sensor, an optical sensor, a sensor having two or more such characteristics, or two or more sensors acting as a unit. As used herein, "sensor" and "control feedback sensor" encompass both a single sensor and two or more communicatively coupled sensors. The tissue processing sensor 215 may further include a chip, which in some embodiments may include a programmable chip. In some embodiments, one or more programmable chips in or connected to the control feedback sensor 215 may be configured to receive and store information sufficient to indicate that a particular cartridge assembly 300 is a new cartridge assembly 300 or a used cartridge assembly 300. In additional or alternative embodiments, the cartridge assembly 300 may further include one or more cartridge assembly programmable sensors (not shown). In such embodiments, the control feedback sensor 215 may be configured to read the cartridge assembly sensor as new or used. In some embodiments, the control feedback sensor may be configured using Bluetooth, Near Field Communication (NFC), I / O, or other communication technologies. 2 The computer may be configured to communicate with one or more computer components via Inter-Integrated Circuit (C) serial communications, a Serial Peripheral Interface (SPI), or any other such communications component.

[0083] 2 also shows the distal end of the pestle shaft (or shaft) 401. In some embodiments, one or more motors (not shown) may actuate the pestle shaft (or shaft) 401 up and down, rotationally, laterally, or two or more such movements in the tissue processing area 202. Thus, in some embodiments, the pestle shaft (or shaft) 401 may extend further into the tissue processing area 202 than is generally shown in FIG.

[0084] 3A, the reader can see an exploded perspective view of a cartridge assembly according to one embodiment of the present disclosure. More specifically, FIG. 3A helps to show cartridge assembly 300, cartridge top cover 301, raised processing area 302, upper processing shelf 303, "A" opening 304, "B" opening 305, "C" opening 306, "D" opening 307, cartridge tab 308, mortar top cap 309, mortar polymer ring 310, mortar cup (or cup) 311, mortar screen (or screen) 312, well plate 313, well A 314, well B 315, well C 316, raised processing element 317, well bottom 317a, well heating cup 318, drive sleeve 319, cartridge bottom cover 320, bottom cover opening 321, docking spindle opening 322, and connection tab 325.

[0085] In some embodiments, cartridge top cover 301 may be generally disk-shaped, as generally shown in FIG. 3A . In other embodiments, cartridge top cover 301 may be square-shaped, spherical-shaped, cubic-shaped, oval-shaped, star-shaped, triangular-shaped, or any other shape capable of implementing the present systems and / or methods. Similarly, in some embodiments, any of "A" opening 304, "B" opening 305, "C" opening 306, and / or "D" opening 307 may be larger or smaller than generally shown in FIG. 3A . Also, in some embodiments, any of "A" opening 304, "B" opening 305, "C" opening 306, and / or "D" opening 307 may be shaped other than circular (e.g., oval, square, diamond, triangular, etc.).

[0086] As seen in FIG. 3A, in some embodiments, one or more of the mortar top cap 309, the mortar polymer ring 310, and / or the mortar cup (or cup) 311 may include one or more wing-shaped tabs on their upper surfaces. As can be inferred from observation of the raised processing area 302, upper processing shelf 303, and "A" opening 304, such wing-shaped tabs may be configured to position the mortar cup (or cup) 311 in an elevated or lowered position within the raised processing area 302, including, by way of example and not limitation, on one or more upper processing shelves 303, or lowered by the "A" opening 304 to a lowered position within the raised processing area 302, including, but not limited to, a mortar screen (or screen) 312, mortar cup (or cup) 311, and a position where a tissue sample is placed in well A 314, well B 315, or well C 316 in the presence of an enzyme solution or buffer solution.

[0087] In some embodiments, the mortar screen (or screen) 312 may include one or more of a membrane filter, a 100 micron filter, a 50 micron filter, a 25 micron filter, a 10 micron filter, a 5 micron filter, a 1 micron filter, a magnetic filter, a microbead filter, a magnetic microbead filter, an antibody / antigen receptor used as a filter, one or more size exclusion columns, one or more centrifuges, or filtering via an ion exchange / polar column. As used herein, the terms "screen" and "filter" are synonymous unless the context indicates otherwise.

[0088] In some embodiments, the well plate 313 may be configured to rotate well A 314, well B 315, or well C 316 directly below the "A" opening 304 and raised processing area 302, such that one or more wells originally located below the "A" opening 304, "B" opening 305, "C" opening 306, or "D" opening 307 may be rotated to align below the "A" opening 304 and raised processing area 302. In some embodiments, such rotation is believed to be achieved when the mortar cup (or cups) 311 is present on the upper processing shelf 303 in the upper position.

[0089] In some embodiments, one or more of well A 314, well B 315, or well C 316 may be pre-filled with a processing medium, including but not limited to, sterile water, a buffer, an enzyme, or a combination thereof.

[0090] Turning to the raised processing element 317, as discussed in more detail elsewhere herein, in some embodiments, with the pestle 400 residing within the mortar cup (or cup) 311 and mortar screen (or screen) 312, one or more motors of the system can drive the pestle shaft (or shaft) 401, causing the pestle head 407 to apply rotational and grinding forces to the mortar screen (or screen) 312 inside the mortar cup (or cup) 311 in the presence of an enzyme solution, a buffer solution, or both, against the tissue sample, with the mortar cup (or cup) 311 and mortar screen (or screen) 312 contacting such fluids in well A 314, well B 315, or well C 316. In some embodiments, the mortar screen (or screen) 312 can reside on the raised processing element 317 within well C 316. In such embodiments, when the pestle 400 applies rotational and grinding forces to the tissue sample against the mortar screen 312 within the mortar cup 311, the raised processing elements 317 may, in some embodiments, provide an additional friction surface that may enhance the mechanical dispersion caused by the rotational and grinding forces of the pestle cap 407. For example, the pestle head 407 may grind one or more tissue sample pieces while one or more raised processing elements 317 hold the tissue sample pieces in place, thereby enhancing the mechanical dispersion of the tissue sample pieces. In some embodiments, the raised processing elements 317 may comprise a separate plate present in one or more of well A 314, well B 315, or well C 316. In some embodiments, the one or more raised processing elements 317 may be configured as one or more linear raised ridges radiating from a central point. In some embodiments, the one or more raised processing elements 317 may be configured as one or more spiral ridges.In alternative embodiments, one or more raised treatment elements 317 may comprise a plurality of tiny pips or dots, which may be randomly arranged, arranged in a grid, arranged in a spiral orientation, or arranged in any geometric pattern.

[0091] In some embodiments, the well bottom 317a may comprise a recess in well C 316. In some embodiments, as seen in FIG. 3A, this may include one or more raised treatment elements 317 on the surface of the well bottom 317a. In some embodiments, the well bottom 317a may be generally curved, flat, conical, or inset. In some embodiments, the shape of the well bottom 317a may be configured to align with the shape of the terminal end of the pestle head 407 and the mortar screen (or screen) 312. As a non-limiting example, if the terminal end of the pestle head 407 is flat, the mortar screen (or screen) 312 may be flat, and the well bottom 317a may be flat or slightly curved. Alternatively, if the terminal end of the pestle head 407 is curved, the mortar screen (or screen) 312 may be curved, and the well bottom 317a may be curved. Similarly, if the pestle head 407 terminates in a conical shape, a conical mortar screen (or screen) 707 may be provided, and the lower well portion 317a may be conical, for example, like the alternative lower well portion 322. If the pestle head 407 terminates in a mating shape, as shown in FIGS. 5I and 5J, for example, a mating mortar cup (or cup) 710 may be provided, and the lower well portion 317a may be mating, for example, like the mating lower well portion 801. In some embodiments, the diameter of the pestle head 407 may be configured to fit within the lower opening of the mortar cup (or cup) 311 or the mating mortar cup (or cup) 710.

[0092] Similarly, as may be disclosed in more detail elsewhere herein, in some embodiments, the diameter of the lower opening of mortar cup (or cup) 311 may be 2 cm. In some embodiments, the diameter of the lower opening of mating mortar cup (or cup) 710 may be 2 cm. Relatedly, in some embodiments, the diameter of pestle head 407 may be 2 cm or slightly less than 2 cm. In related embodiments, the diameter of well lower portion 317 a may be 2 cm or slightly more than 2 cm. In some embodiments, the diameter of mating well lower portion 801 may be 2 cm or slightly more than 2 cm. In some embodiments, the diameter of mortar screen (or screen) 312 may be 2 cm or slightly more than 2 cm.

[0093] Similarly, in some embodiments, the diameter of the lower opening of mortar cup (or cup) 311 may be 1 cm. In some embodiments, the diameter of the lower opening of mating mortar cup (or cup) 710 may be 1 cm. Relatedly, in some embodiments, the diameter of pestle head 407 may be 1 cm or slightly less than 1 cm. In related embodiments, the diameter of lower well portion 317a may be 1 cm or slightly more than 1 cm. In some embodiments, the diameter of lower mating well portion 810 may be 1 cm or slightly more than 1 cm. In some embodiments, the diameter of mortar screen (or screen) 312 may be 1 cm or slightly more than 1 cm.

[0094] In relation to the above, in some embodiments, the diameter of the lower opening of mortar cup (or cup) 311 may be 2 cm to 5 cm. In some embodiments, the diameter of the lower opening of mating mortar cup (or cup) 710 may be 2 cm to 5 cm. In this regard, in some embodiments, the diameter of pestle head 407 may be 2 cm to 5 cm. In related embodiments, the diameter of well lower portion 317a may be 2 cm to 5 cm. In some embodiments, the diameter of mating well lower portion 810 may be 2 cm to 5 cm. In some embodiments, the diameter of mortar screen (or screen) 312 may be 2 cm to 5 cm.

[0095] Similarly, in some embodiments, the diameter of the lower opening of mortar cup (or cup) 311 may be greater than 5 cm. In some embodiments, the diameter of the lower opening of mating mortar cup (or cup) 710 may be greater than 5 cm. Relatedly, in some embodiments, the diameter of pestle head 407 may be greater than 5 cm. In related embodiments, the diameter of well bottom 317a may be greater than 5 cm. In some embodiments, the diameter of mating well bottom 810 may be greater than 5 cm. In some embodiments, the diameter of mortar screen (or screen) 312 may be greater than 5 cm.

[0096] Similarly, in some embodiments, the diameter of the lower opening of mortar cup (or cup) 311 may be less than 1 cm. In some embodiments, the diameter of the lower opening of mating mortar cup (or cup) 710 may be less than 1 cm. Relatedly, in some embodiments, the diameter of pestle head 407 may be less than 1 cm. In related embodiments, the diameter of lower well portion 317a may be less than 1 cm. In some embodiments, the diameter of lower mating well portion 810 may be less than 1 cm. In some embodiments, the diameter of mortar screen (or screen) 312 may be less than 1 cm.

[0097] As generally seen in FIG. 3A , one or more of well A 314, well B 315, or well C 316 may be shaped generally as a cylinder, with at least one sidewall and at least one distal or bottom surface. In some embodiments, one or more of well A 314, well B 315, or well C 316 could have a shape other than a cylinder, such as a sphere, a rectangular prism, or any other geometric shape. Also, at least one raised treatment element may be disposed along one or more of at least one sidewall or bottom distal end of one or more of well A 314, well B 315, or well C 316.

[0098] It is contemplated that in some embodiments, well C316 may have a narrower or wider diameter and a deeper or shallower vertical length to accommodate differences in buffer solutions and in swelling ratios of the regenerative epithelial solutions of the present disclosure. As a non-limiting example, in some embodiments, well C316 may have a narrower, shallower, or both a narrower and shallower diameter to accommodate processing of smaller volumes of buffer (e.g., 5 ml) or larger volumes of buffer (e.g., 500 ml) by a system for automated preparation of a regenerative epithelial suspension according to at least one embodiment of the present disclosure.

[0099] 2 and 3A , in some embodiments, the cartridge bottom cover 320, bottom cover opening 321, and docking spindle opening 322 may be configured to encase the heating element 205, docking spindle 206, and control feedback sensor 215. In other embodiments, the cartridge bottom cover 320 may be configured to overlie one or more of the heating element 205, docking spindle 206, and control feedback sensor 215 with no cutouts. It is contemplated that in some embodiments, one or more motors in the housing 201 may drive the docking spindle 206, thereby rotating the docking spindle 206 relative to the drive sleeve 319 and subsequently rotating the well plate 313 within the cartridge top cover 301.

[0100] In some embodiments, cartridge assembly 300 and its components (including, but not limited to, cartridge top cover 301, raised treated area 302, "A" opening 304, "B" opening 305, "C" opening 306, "D" opening 307, well plate 313, well A 314, well B 315, well C 316, heating cup 318, drive sleeve 319, and cartridge bottom cover 320) may be configured in a linear fashion. By way of non-limiting example, in some embodiments, cartridge top cover 301 may be generally rectangular in shape, with raised treated area 302 and its "A" opening 304, "B" opening 305, "C" opening 306, and "D" opening 307 disposed in a line along the top edge of cartridge top cover 301. Correspondingly, in such an embodiment, by way of non-limiting example, well plate 313 may also be generally rectangular, with well A 314, well B 315, and well C 316 arranged in a line. In such an embodiment, cartridge bottom cover 320 may also be configured in a rectangular shape. Of course, in the above example, the linear arrangement of elements such as raised treated area 302, "A" opening 304, "B" opening 305, "C" opening 306, "D" opening 307, well A 314, well B 315, and well C 316, heating cup 318, or drive sleeve 319 does not necessarily require a rectangular configuration of elements (including, but not limited to, cartridge top cover 301, well plate 313, and cartridge bottom cover 320). Rather, the cartridge top cover 301, well plate 313, and cartridge bottom cover 320, as well as any other elements discussed herein, may be configured in different shapes, such as oval, square, or circular, while still being configured to accommodate linear motion and processing sequences.

[0101] Those skilled in the art will appreciate that in alternative embodiments, one or more of "A" opening 304, "B" opening 305, "C" opening 306, "D" opening 307, well A 314, well B 315, or well C 316 may be labeled with alternative identifying marks, such as numbers, symbols, colors, color coding, or other sequential or non-sequential identifying marks. As a non-limiting example, in some embodiments, "A" opening 304, "B" opening 305, "C" opening 306, and "D" opening 307 may be numbered 1, 2, 3, and 4, while well A 314, well B 315, and well C 316 may have their respective alphabetical designations. Alternatively, for purposes of example only, in some embodiments, "A" opening 304, "B" opening 305, "C" opening 306, and "D" opening 307 may have their respective alphabetical designations, while wells A 314, B 315, and C 316 may be numbered 1, 2, 3, and 4.

[0102] In some embodiments, tissue samples may be collected by a corkscrew slicer, such as a food processor, electroporation, biopsy, punch biopsy, dermatome, DermaBlade®, or any other device or method known in the relevant art for collecting skin and / or tissue samples.

[0103] In some embodiments, enzyme inactivation may be achieved in one or more of wells A314, B315, or C316 by washing the tissue sample in a buffer, one or more trypsin inhibitors, vacuum removal, semi-permeable membranes, enzyme specific receptors in a column filtering process, pressure differentials, magnetic microbeads, or any other method known in the relevant art.

[0104] In some embodiments, fluid movement within cartridge assembly 300 may be achieved by syringe plunger / pulling action, magnetic fluid movement, powered fluid movement (e.g., a stepper), vacuum pressure, fluid pressure, gravity, concentration gradient, or any other such mechanism known in the art.

[0105] In some embodiments, the regenerative epithelial suspension may be applied to a patient by spraying the treatment area directly with a spray nozzle, spraying the treatment area directly with an ultrasonic or piezoelectric atomizer, spraying the treatment area directly with a rotary spray nozzle with a variable spray pattern, dripping onto the treatment area with a dripper or syringe, applying to a dressing or vehicle, combining the regenerative epithelial suspension with a foam, combining the regenerative epithelial suspension with an electrospun bandage, combining the regenerative epithelial suspension with clothing (such as, but not limited to, gloves or socks), combining the regenerative epithelial suspension with elements necessary to make a paste, combining the regenerative epithelial suspension with elements necessary to make an ointment, combining the regenerative epithelial suspension with elements necessary to make a balm, combining the regenerative epithelial suspension with elements necessary to make a "lipstick" style applicator, or any other application method or combination known in the art.

[0106] Continuing to refer to FIG. 3A , a viewer can see that in some embodiments, well C 316 can include an oval-shaped upper well portion, one or more moon-shaped lower well shelves, a lower well portion along which, in some embodiments, a raised processing element 317 is disposed, and an interior well portion disposed in the center of the lower well portion. As seen generally in FIG. 3A , in some embodiments, the lower well portion can include a shallow fluid receptacle. Also, as seen in FIG. 3A , in some embodiments, the lower well portion can have a generally circular shape. A viewer of FIG. 3A can also see that the interior well portion itself can be circular and can be disposed in the center of the lower well portion, as described in some embodiments.

[0107] Continuing to focus on FIG. 3A , one skilled in the art will recognize that the well bottom, along with the raised processing elements 317, may slope at various angles toward the well interior. While FIG. 3A discloses a relatively flat slope, other embodiments may provide a steeper well interior, as seen in FIG. 7 . In some embodiments, the well interior may be shaped as a shallow tray with a slight slope toward its center, as generally disclosed in FIG. 3A , or may be more conical, as shown in FIG. 7 . One skilled in the art will appreciate that in alternative well bottom embodiments such as those described above, the raised processing elements 317 may include more or fewer elements than those shown in FIG. 3A , or may have a different orientation than shown. Additionally, in some embodiments, the well interior may include flat surfaces, may itself include depressions, or may include raised elements in some embodiments.

[0108] 3B shows additional details regarding, among other elements, the cartridge top cover 301, the raised processing area 302, and the upper processing shelf 303. As seen in FIG. 3B, the raised processing area 302 may generally include one or more vertical elements 323, each of which may further include a cutout 324. As can be seen with reference to FIG. 7A, the raised processing area 302 may be configured such that the cutouts 324 may be configured to allow passage of one or more mortar cup (or cup) tabs 701 therethrough, and the upper processing shelf 303 may be configured such that the mortar cup (or cup) tabs 701 may reside on a surface of the upper processing shelf to form a space below the mortar cup (or cup) 311 and the lower portion of the raised processing area 302. In some embodiments, such a space allows the well plate 313 to rotate one or more of well A 314, well B 315, or well C 316 directly below the mortar cup (or cup) 311. Conversely, the system may be configured such that rotation of the pestle 400 removably engaged with the mortar cup (or cup) 311 can twist the mortar cup (or cup) tab 701 off the upper processing shelf 303 and through the notch 324, thereby lowering the mortar cup (or cup) 311 to a lower position within the vertical element 323, and at least a portion of the mortar cup (or cup) 311 may be inside well A 314, well B 315, or well C 316 at the lower position within the raised processing area 302.

[0109] 3C generally discloses a bottom perspective view of cartridge top cover 301. In FIG. 3C, the viewer can see connection tab notches 326 and connection tab lock openings 327, each of which, in some embodiments, can be configured to receive a connection tab 325. In some embodiments, during assembly, each connection tab 325 slides through a connection tab notch, and an end of each connection tab 325 extends through a connection tab lock opening 327, thereby securing cartridge bottom cover 320 to cartridge top cover 301.

[0110] 3D generally discloses a top perspective view of well plate 313, along with well A 314, well B 315, and well C 316. In FIG. 3D, the viewer can also see component opening 328, raised treated area cutout 329, and tab stopper 330.

[0111] 3E generally discloses an alternative well plate 331, an alternative well bottom 332, and an alternative well interior 333. In some embodiments, including but not limited to, those configured to prepare cell suspensions for small area treatment, the present invention may provide smaller wells A 314, B 315, and C 316, as well as more conical well bottoms, such as alternative well bottom 332. Also, in some embodiments, the circumference of alternative well bottom 332 may be smaller than the circumference of the well bottom shown in FIG. 3A. This may allow for effective processing and collection of smaller skin samples or smaller fluid volumes, as a non-limiting example, due to the steeper slope and smaller size of alternative well bottom 332.

[0112] FIG. 3F generally discloses the underside of one embodiment of well plate 313, along with well A 314, well B 315, and well C 316, as well as component opening 328 and raised treated area cutout 329 on the underside.

[0113] FIG. 3G generally discloses a top perspective view of cartridge bottom cover 320, connection tabs 323, bottom cover opening 321, and docking spindle opening 322. While a viewer can see that in some embodiments, cartridge bottom cover 320 includes a plurality of raised elements radiating outward from cartridge bottom cover 320, as generally disclosed in FIG. 3G, such a configuration is shown in FIG. 3G for illustrative purposes only, and other embodiments are contemplated, such as a smooth surface on the top surface of cartridge bottom cover 320, a grid pattern on the top surface of cartridge bottom cover 320, etc. In some embodiments, the smooth surfaces on the top surfaces of cartridge bottom cover 320 and other associated elements of cartridge 300 (e.g., cartridge top cover 301 and well plate 313) may have alternative shapes, such as a square or rectangle.

[0114] FIG. 3H generally discloses a bottom perspective view of cartridge bottom cover 320, connection tabs 323, bottom cover opening 321, and docking spindle opening 322.

[0115] With reference to Figure 4, the viewer can appreciate an exploded perspective view of a pestle 400, in accordance with one embodiment of the present disclosure. More specifically, Figure 4 helps disclose the pestle 400, pestle shaft (or shaft) 401, pestle top cap 402, pestle spring 403, pestle spring cap 404, pestle bottom cap 405, pestle body 406, pestle head 407, first pestle retention wire 408, second pestle retention wire 409, pestle body ridge 410, and pestle head internal notch 411.

[0116] In some embodiments, the pestle 400 may comprise a pestle top cap 402, a pestle spring 403, a pestle spring cap 404, a pestle bottom cap 405, a pestle body 406, a pestle head 407, a first pestle retaining wire 408, and a second pestle retainer 409. In such embodiments, the pestle shaft (or shaft) 401 may comprise certain elements of the base unit 200, and the pestle 400 may comprise other elements shown in FIG.

[0117] However, in some embodiments, the pestle 400 may include all of the pestle shaft (or shaft) 401, the pestle top cap 402, the pestle spring 403, the pestle spring cap 404, the pestle bottom cap 405, the pestle body 406, the pestle head 407, the first pestle holding wire 408, and the second pestle holding wire 409.

[0118] 4 may not be present in alternative embodiments of the pestle 400. For example, by way of non-limiting example, in some embodiments, the pestle 400 may simply include a pestle top cap 402, a pestle body 406, and a pestle head 407. In some embodiments, the pestle 400 may simply include a pestle top cap 402 and a pestle head 407. In other embodiments, the pestle 400 may include an alternative pestle head 407 with a closed top and a notch in the closed top configured to receive the pestle shaft (or shaft) 401.

[0119] Similarly, in some embodiments, the pestle 400 may include a pestle top cap 402, a pestle spring 403, a pestle spring cap 404, a pestle bottom cap 405, a pestle body 406, and a pestle head 407, but not a first pestle holding wire 408 and a second pestle holding wire 409. Similarly, in some embodiments, the pestle 400 may include a pestle top cap 402, a pestle spring cap 404, a pestle bottom cap 405, a pestle body 406, and a pestle head 407, but not a pestle spring 403, a first pestle holding wire 408, and a second pestle holding wire 409.

[0120] In some embodiments, the pestle spring 403 may instead include a force sensor, transducer element, force transducer, force measurement sensor, load sensor, load cell, tensile force sensor, compressive force sensor, tension sensor, load pin, load bearing, stress sensor, customized sensor, pressure sensor, or another equivalent element.

[0121] In some embodiments, the pestle 400 may apply rotational and / or grinding forces to the tissue sample within the mortar cup (or cup) 311 against the sidewalls of the mortar cup (or cup) 311 as well as the mortar screen (or screen) 312. As seen in more detail in FIG. 5, the pestle head 407 may be configured to provide a friction surface that transfers these rotational and grinding forces to the tissue sample. However, in other embodiments, the pestle 400 may be configured with different structures suitable for alternative tissue dispersion methods. In some embodiments, such tissue disaggregation may be accomplished by, but is not limited to, ultrasonic disaggregation, piezoelectric disaggregation, rotational disaggregation with one or more blades, linear disaggregation with one or more blades, mixing devices, punch-through disaggregation, punch-through disaggregation with a mesh or filter, multi-stage punch-through disaggregation, gratings such as those with blade surfaces in the shape of a cheese grater, "slap chop" disaggregation, spiral grinding disaggregation (such as, but not limited to, a pestle with raised spiral grooves configured to grind the tissue against a mortar wall), string slicer disaggregation, shaver disaggregation, actuation disaggregation (such as, but not limited to, actuation in the presence of microbeads), microbubble or cavitation disaggregation, magnetic disaggregation, and electromagnetic disaggregation. In some embodiments, the pestle 400 may be configured differently or may be replaced with another assembly suitable for such alternative disaggregation means.

[0122] In some embodiments, the pestle shaft (or shaft) 401 may be adapted to mate with the pestle top cap 402 in a key / lock arrangement as generally shown in Figure 4. However, in other embodiments, by way of non-limiting example, the pestle shaft (or shaft) 401 may be adapted to mate with the pestle top cap 402 via a magnetic connection engagement, a luer lock, a snap fit engagement, a bayonet style engagement, an air hose quick connect engagement, a press fit engagement, a "Mix2Vial" engagement, and any other form of temporary, reversible, and in some embodiments, permanent engagement known in the relevant art. In this regard, the pestle shaft (or shaft) 401 may generally have the keyed shape disclosed in Figure 4, or may be shorter, longer, wider, narrower, or may have one or more protrusions useful for connecting with the pestle top cap 402. In some embodiments, the pestle shaft (or shaft) 401 may include components of the housing 201 rather than elements of the pestle 400 .

[0123] In some embodiments, the pestle shaft (or shaft) 401, when inserted through the pestle top cap 402, may compress the pestle spring 403 against the pestle spring cap 404. This force then causes the pestle spring cap 404 to contact the pestle bottom cap 405, which in turn contacts the pestle body 406, which in some embodiments may be in releasable mating contact with the pestle head 407. Thus, in some embodiments, one or more motors contained within the housing 201 may rotate, oscillate, actuate, or otherwise manipulate the pestle shaft (or shaft) 401, thereby rotating, oscillating, actuating, or otherwise manipulating the pestle 400.

[0124] For purposes of the above example, in some embodiments, one or more motors housed in the housing 201 may rotate the pestle shaft (or shaft) 401 such that one or more raised protrusions on one or more sides of the pestle shaft (or shaft) 401 may come into contact with one or more elements on the underside of the pestle top cap 402. In such embodiments, when the pestle shaft (or shaft) 401 comes into contact with the pestle top cap 402 in this manner, the pestle top cap 402 may rotate the pestle body 406, which in turn may rotate the pestle head 407. Thus, in such embodiments, when the one or more motors rotate the pestle shaft (or shaft) 401 within the pestle 400, the pestle head 407 generates a rotational force which, if the pestle head 407 is in contact with a tissue sample, may be applied to the tissue sample.

[0125] Also, in some embodiments, the sequence described above may involve one or more motors within the housing 201 configured to press the pestle shaft (or shaft) 401 against the pestle spring 403, which presses the pestle spring cap 404, which presses the pestle bottom cap 405, which will come into contact with the pestle body 406, which itself may already be in engaging contact with the pestle head 407. In embodiments, if the pestle head 407 is in contact with the tissue sample, the sequence described above may cause the pestle head 407 to apply pressure against the tissue sample, which in some embodiments may be disposed along the mortar screen (or screen) 312. In some embodiments, the force constant of the pestle spring 403 may be adjusted to apply a particular spring force in the pestle spring 403 axis through the pestle body 406 and the pestle head 407. Such pressure on the tissue sample can impart a grinding force to the tissue, and when applied in combination with one or more rotational movements by the pestle head 407 on the tissue sample, can apply both a rotational force and a grinding force to the tissue sample.

[0126] In some embodiments, the above-described sequence in which one or more motors contained within the housing 201 rotate the pestle head 401, causing the pestle shaft (or shaft) 401 to contact one or more elements on the underside of the pestle top cap 402, which in turn rotates the pestle top cap 402, which in turn rotates the pestle body 406, which in turn rotates the pestle head 407, may be useful for raising or lowering the mortar cup (or cup) 311 within the raised treatment area 302. In some embodiments, by way of non-limiting example, a clockwise rotational movement of the pestle shaft (or shaft) 401 relative to the pestle top cap 402 raises the mortar cup (or cup) 311 and pestle 400 onto the upper processing area 303, thereby positioning the mortar cup (or cup) 311 and pestle 400 onto the upper processing area 303 via the mortar top cap 309, the mortar polymer ring 310, or one or more tabs on the mortar cup (or cup) 311, which in some embodiments allows the well plate 313 to rotate well A 314, well B 315, and well C 316 directly below the raised processing area 302. In some embodiments, a counterclockwise rotational movement of the pestle shaft (or shaft) 401 relative to the pestle top cap 402 raises the mortar cup (or cup) 311 and pestle 400 onto the upper processing area 303, and the mortar cup (or cup) 311 and pestle 400 are positioned onto the upper processing area 303 by the mortar top cap 309, the mortar polymer ring 310, or one or more tabs on the mortar cup (or cup) 311, which in some embodiments allows the well plate 313 to rotate well A 314, well B 315, and well C 316 directly below the raised processing area 302.

[0127] In some embodiments, a clockwise rotational motion by the pestle shaft (or shaft) 401 relative to the pestle top cap 402 may lower the mortar cup (or cup) 311 and pestle 400 from the upper processing area 303 into one or more of well A 314, well B 315, or well C 316 to perform one or more tissue processing steps. In some embodiments, a counterclockwise rotational motion by the pestle shaft (or shaft) 401 relative to the pestle top cap 402 may lower the mortar cup (or cup) 311 and pestle 400 from the upper processing area 303 into one or more of well A 314, well B 315, or well C 316 to perform one or more tissue processing steps. In certain preferred embodiments, the rotational motion applied by the pestle shaft (or shaft) 401 relative to the pestle top cap 402 is counterclockwise.

[0128] As seen in FIG. 4 , in some embodiments, the pestle 400 may include four pestle head internal notches 411, which in some embodiments may correspond to four pestle body ridges 410 disposed on the pestle body 406. In some embodiments, the pestle 400 may include three pestle head internal notches 411, which in some embodiments may correspond to three pestle body ridges 410. In some embodiments, the pestle 400 may include two pestle head internal notches 411, which in some embodiments may correspond to two pestle body ridges 410. In some embodiments, the pestle 400 may include one pestle head internal notch 411, which in some embodiments may correspond to one pestle body ridge 410. In some embodiments, the pestle 400 may include five or more pestle head internal notches 411, which in some embodiments may correspond to five or more pestle body ridges 410.

[0129] 5A-5H illustrate a series of alternative embodiments of a pestle head 407, according to various embodiments of the present disclosure. More specifically, FIG. 5A serves to illustrate a pestle head 407 having a smooth exterior and a terminal ridge or "bump." FIG. 5B serves to illustrate a pestle head 407 having a smooth exterior and a terminal ridge spiral shape.

[0130] FIG. 5C illustrates a pestle head 407 having an external raised spiral pattern and a terminal raised spiral shape. FIG. 5D illustrates a pestle head 407 having an external raised spiral pattern and terminal raised ridges or "bumps." FIG. 5E illustrates a pestle head 407 having a smooth external and terminal raised lattice pattern. FIG. 5F illustrates a pestle head 407 having an external structure consisting of repeating right angles and a terminal raised radial pattern. In some embodiments, the pestle head 407 may be constructed from one or more rubbers, including, but not limited to, silicone rubber, natural rubber, butadiene rubber, butyl rubber (IIR), ethylene propylene diene monomer (EPM or EPDM), fluoroelastomer (FKM / Viton), isoprene rubber, nitrile rubber (NBR), or styrene butadiene rubber. Alternatively, the pestle head 407 may be constructed from a metal, including, but not limited to, stainless steel, copper, titanium, cobalt chromium, aluminum, magnesium, gold, platinum, silver, iridium, tantalum, etc. Alternatively, it may be constructed from any type of plastic, including, but not limited to, polycarbonate, polypropylene, acrylonitrile butadiene styrene (ABS), polyethylene, polymethyl methacrylate, polyvinyl chloride (PVC), polyethylene terephthalate glycol (PETG), polylactic acid (PLA), or any combination of such materials or other types of rubber, metal, or plastic known in the art. In some embodiments, the pestle head 407 may be removable and replaceable.

[0131] 5G and 5H also generally disclose raised side elements 501 and raised element openings 502, as well as wide tapered pestle head 503, narrow tapered pestle head 505, wide pestle head point 504, narrow pestle head point 506, wide spiral raised groove pattern 505, and narrow spiral raised groove pattern 509. It is noted that raised side elements 501 and raised element openings 502 are also seen in FIGS. 5A-5E.

[0132] In some embodiments, the wide tapered pestle head 504 or the narrow tapered pestle head 505 may be paired with a tapered mortar cup (or cup) 801, as seen in FIG. 8B. In this regard, the wide tapered pestle head 504 or the narrow tapered pestle head 505 may be configured to be used in conjunction with an alternative well lower portion 702, such as generally disclosed in FIG. 7A, integral with the tapered mortar cup (or cup) 801. Furthermore, in some embodiments, the wide tapered pestle head 504 or the narrow tapered pestle head 505 may include any of the raised elements disclosed in FIGS. 5A-5F in addition to or in combination with the wide spiral raised groove pattern 508 or the narrow spiral raised groove pattern 509.

[0133] 5A-5E, 5G, and 5H, in some embodiments, two raised side elements 501 may generally form a ring around the pestle head 407, with the rings separated by one or more raised element openings 502. In some embodiments, the ring shape may include a single-opening ring, with a single raised element opening 502 disposed along a single raised side element 501. In some embodiments, a double-opening ring shape may include two raised side elements 501 separated by two raised element openings 502. In some embodiments, the ring shape may include three or more raised side elements 501 separated by three or more raised element openings 502.

[0134] In some embodiments, two or more raised side elements 501 may be disposed in any order along the pestle head 407, including, but not limited to, an alternating order. Thus, by way of example, in a two-ring configuration, one or more raised side elements 501 may be disposed one above the other along the pestle head 407, separated by one or more raised element openings 502. In some such embodiments, the raised element openings 502 may be aligned with each other relative to the longitudinal axis of the pestle head 407, or in some embodiments, may be staggered or in any order relative to each other and the longitudinal axis of the pestle head 407.

[0135] Those skilled in the art will also appreciate that in some embodiments, the raised side elements 501 may be wider, narrower, taller, or shorter than those shown in Figures 5A-5E, 5G, and 5H. Also, in some embodiments, the raised element openings 502 may not be tapered. In variations including two or more raised element openings 502, some raised element openings 502 may be tapered while others are not tapered. In embodiments including two or more raised element openings 502, the raised element openings may be vertically aligned along the pestle cap 407, staggered, or misaligned relative to the pestle cap 407, or some raised element openings 502 may be aligned with one another and some may be misaligned relative to the pestle cap 407.

[0136] In some embodiments, such as those generally shown in Figures 5A-5E, 5G, and 5H, two or more raised side elements 501 may have one or more openings. In some such embodiments (such as the exemplary, non-limiting embodiments generally disclosed in Figures 5A-5E, 5G, and 5H), the raised ring element may taper to a point on either side of the opening or openings. In a preferred embodiment, the raised element may comprise a raised ring including two openings, and the raised ring element may taper to a point on either side of the opening or openings.

[0137] Furthermore, in some embodiments, the raised side elements, generally disclosed in Figures 5A-5E, 5G, and 5H, which in some embodiments may further include one or more openings, may be configured such that when the pestle head 407 is inserted into the mortar cup (or cup) 311 beyond the internal ridge 803, friction between the pestle head 407 and the interior of the mortar cup (or cup) 311 may cause the pestle 400 to raise or otherwise manipulate the mortar cup (or cup) 311, including, but not limited to, lowering the mortar cup (or cup) 311 at a controlled or intended speed, rotating the mortar cup (or cup) 311, or actuating or moving the mortar cup (or cup) 311.

[0138] 51 generally discloses a mating pestle head 517 having a mating end 509 with an externally raised helical element 511. In such embodiments, the mating end 509 may comprise a cylinder having a smaller diameter than the pestle head cylinder 508. In some embodiments, the transition end 501 between the mating end 509 and the pestle head cylinder 508 may comprise a beveled or curved edge.

[0139] 5I also generally discloses different angles on the raised side element 501. As seen in FIG. 5I, the raised side element 501 may include a flat upper edge 514 and a tapered lower edge 515. In some embodiments, the raised side element 501 may be configured to allow the pestle head 407 to enter the interior of the mortar cup (or cup) 311 but prevent it from exiting the interior of the mortar cup (or cup) 311. In such an embodiment, the tapered lower edge 515 allows the pestle head 407 to push the mortar top cap 309, the mortar polymer ring 310, or both the mortar top cap 309 and the mortar polymer ring 310 through which it passes and enter the interior of the mortar cup (or cup) 311, while the flat upper edge 514 prevents the pestle head 407 from exiting the interior of the mortar cup (or cup) 311. 5I also generally discloses the raised helical element 511 and helical gap 512 along with the mating end distal edge 513. In some embodiments, the raised side element 501 and its flat upper edge 514 and / or tapered lower edge 515 may perform the same function for the wide tapered pestle cap head 503, the narrow tapered pestle cap head 505, the mating pestle head 517, the conical pestle head, and any other pestle head described or discussed in this disclosure. 8A, 8B, and elsewhere herein, when the mating pestle head 517 rotates within a corresponding mating mortar cup (or cup) 710, the raised helical element 511 may grind tissue against the interior sidewall of the mating mortar cup (or cup) 710. In some embodiments, the tissue in such solution may be allowed to collect in one or more helical gaps 512 or circulate within the mating mortar cup (or cup) 710.

[0140] Figure 5J generally discloses one embodiment of a mating pestle head 517 having a mating end 509 and a plurality of protrusions 516 disposed along the distal end of the mating end 509. While in some embodiments, the exterior of the mating end 509 may be generally smooth as shown in Figure 5J, in alternative embodiments, the exterior of the mating end 509 may include raised helical elements 511 and corresponding helical gaps 512, additional protrusions, vertically raised elements, horizontally raised elements, diagonally raised elements, recesses, outwardly extending elements, and other such elements.

[0141] 6 is a flowchart useful in illustrating a process sequence of a system for automated preparation of a regenerative epithelial suspension, according to at least one embodiment of the present disclosure. In some embodiments, one or more elements of the process sequence disclosed in FIG. 6 or one or more sequences described elsewhere herein may be omitted or performed in a different order.

[0142] After powering on the system for automated preparation of regenerative epithelial suspension and initiating a processing sequence, the system may first engage the pestle with the mortar cup (or cup) (601). The system may then cause the pestle to move the mortar cup (or cup) from the upper processing shelf to Well A (602). The heating element may then warm the enzyme solution in Well A (603). The system may then cause the pestle shaft (or shaft) to actuate the pestle so that the pestle contacts the tissue sample with the enzyme solution (604). The system may then cause the pestle to raise the mortar cup (or cup) to an "up" position (605). The system may then press the pestle against the tissue sample three times to squeeze the enzyme solution out of the tissue sample (606). The system may then rotate the well plate so that Well B is positioned directly below the mortar cup (or cup) (607). The system may then cause the pestle to move the mortar cup (or cup) from the upper processing shelf to well B (608). The system may then cause the pestle shaft (or shaft) to actuate the pestle so that the pestle contacts the tissue sample with a first amount of buffer solution (609). The system may then cause the pestle to raise the mortar cup (or cup) to an "up" position (610). The system may then press the pestle against the tissue sample three times to squeeze the buffer solution out of the tissue sample (611). The system may then rotate the well plate so that well C is positioned directly below the mortar cup (or cup) (612). The system may then cause the pestle to move the mortar cup (or cup) from the upper processing shelf to well C (613). The system may then actuate the pestle shaft (or shafts) to apply rotational and grinding forces to the tissue sample in the presence of the second amount of buffer solution, the mortar screen (or screens), and the raised processing elements (614).The system may then cause the pestle to raise the mortar cup (or cups) to the "up" position (615). The system may then press the pestle against the tissue sample three times to squeeze the buffer solution out of the tissue sample (616). The system may then rotate the well plate so that well C is directly below opening D in the cartridge top cover (617). The sensor may then send a signal to a memory unit to record the just-completed processing event (618).

[0143] In some embodiments, in step 614, the system may alternatively cause the pestle shaft (or shaft) to actuate the pestle to apply a linear force to the tissue sample in addition to or instead of the rotational and grinding forces in the presence of the second amount of buffer solution, the mortar screen (or screens), and the raised processing element (614). Also, in some embodiments, in step 614, the system may cause the pestle to apply a linear force, rotational force, or grinding force, or a combination thereof, to the tissue sample without the presence of the mortar screen (or screens). Similarly, in some embodiments, in step 614, the system may cause the pestle to apply a linear force, rotational force, or grinding force, or a combination thereof, to the tissue sample without the presence of the raised processing element.

[0144] Also, in some embodiments, the names of the wells and openings listed in the processing sequence disclosed in FIG. 6 may include one or more numbers, one or more symbols, one or more raised features, or a combination thereof.

[0145] 7A and 7B generally disclose an embodiment of a mortar cup (or cup) according to an embodiment of the present disclosure. More specifically, with respect to Fig. 7A, attention is paid to tab 701, tab post 702, internal ridge 703, mortar top 704, mortar bottom 705, and inter-tab notch 706. With respect to Fig. 7B, attention is paid to conical mortar screen (or screen) 707, tapered mortar cup (or cup) design 708, and mortar cup (or cup) ring 709.

[0146] 7A, the upper mortar portion 704 may be wider in diameter than the lower mortar portion 705, and the internal ridge 703 may form a transition surface between the upper mortar portion 704 and the lower mortar portion 705. As described elsewhere, in some embodiments, the pestle 400 may be inserted into the mortar cup (or cup) 311 such that the raised side elements 501 on the pestle head 407 pass through the upper mortar portion 704 and press across the internal ridge 703 to frictionally engage the lower mortar portion 705. In such embodiments, the pressure and friction from the raised side elements 501 against the surface of the lower mortar portion 705 may enable the pestle to lift the mortar cup (or cup) 311 during one or more processing events or steps disclosed herein. In other embodiments, the raised side element 501 may pass through the mortar top 704, push across the internal ridge 703, and press against the mortar bottom 705, while being prevented from exiting the mortar cup (or cup) 311 by the mortar top cap 309, the mortar polymer ring 310, or both the mortar top cap 309 and the mortar polymer ring 310.

[0147] FIG. 7C is a side view of a mating mortar cup (or cup) 710.

[0148] Figure 8A generally discloses one embodiment of a pestle, mortar cup (or cup), and well C combination including a pestle 400 with a mating pestle head 517, a mating mortar cup (or cup) 710, and a mating well C802 with a mating well lower portion 801. Figure 8B generally discloses an alternative embodiment of a pestle, mortar cup (or cup), and well C combination including a pestle 400 with a mating pestle head 517, a mating mortar cup (or cup) 710, and a mating well C802 with a mating well lower portion 801.

[0149] FIG. 9A is a top view of mating well C 802 having mating well bottom 801 along with a tall raised processing element 901.

[0150] FIG. 9B is a top view of mating well C 802 having mating well bottom portion 801 along with a low height raised processing element 902.

[0151] FIG. 9C is a side perspective view of mating well C802 and lateral protrusion 903.

[0152] FIG. 10A generally discloses a perspective view of one embodiment of a four-piece type of packaging including two clamshell tool set pairs, a set of tools, and a bottom tray configured to receive the set of tools. In embodiments, the present disclosure may provide two clamshell pairs, each of which may include a negative half of the tool configured as a recess for receiving the tool. The tool is intended for use as part of a system for automated cell suspension preparation or as an adjunct to a system for automated cell suspension preparation, and each part may include a mirror image of the other. Thus, each integrated clamshell pair may be configured to removably contain one or more tools. As seen in FIG. 10A , in some embodiments, the present disclosure may include two clamshell pairs. In other embodiments, a system for automated cell suspension preparation may include one clamshell pair containing tools, two clamshell pairs containing tools, three clamshell pairs containing tools, or four or more clamshell pairs containing tools.

[0153] It is contemplated that in some embodiments, one or more tools configured for use in a system for automated preparation of cell suspensions may be included in trays having a shape other than the clamshell pair configuration shown in FIG. 10A. By way of non-limiting example, in some embodiments, the tools may be included in a single tray, two trays, or three or more trays. The one or more trays include one or more recesses configured to receive one side of each tool and a removable protective cover disposed along the surface of the tray.

[0154] In the embodiment generally disclosed in FIG. 10A , the bottom tray is shown as an open-topped pan with a single corner notch design and an upper lip disposed along the entire opening. However, in other embodiments, the bottom tray may comprise a box, bag, sphere, or any other shaped object capable of containing a tool. Similarly, in some embodiments, the tray element and one or more clamshell companion pieces may include zero, two, or more corner notch elements.

[0155] FIG. 10B is a top view of the upper clamshell component of the first clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10B may be configured to receive one or more scalpels, one or more syringes, one or more needles, and one or more vials. In some embodiments, the recesses of the clamshell component shown in FIG. 10B may be configured to contain two or more sizes of tools, such as a small, medium, or large scalpel, a small, medium, or large syringe, a small, medium, or large needle, and a small, medium, or large vial. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10B.

[0156] FIG. 10C is a bottom view of the upper clamshell component of the first clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10C may be configured to receive one or more scalpels, one or more syringes, one or more needles, and one or more vials. In some embodiments, the recesses of the clamshell component shown in FIG. 10C may be configured to contain two or more sizes of tools, such as a small, medium, or large scalpel, a small, medium, or large syringe, a small, medium, or large needle, and a small, medium, or large vial. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10C.

[0157] FIG. 10D is a top view of the lower clamshell component of the first clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10D may be configured to receive one or more scalpels, one or more syringes, one or more needles, and one or more vials. In some embodiments, the recesses of the clamshell component shown in FIG. 10D may be configured to contain two or more sizes of tools, such as a small, medium, or large scalpel, a small, medium, or large syringe, a small, medium, or large needle, and a small, medium, or large vial. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10D.

[0158] FIG. 10E is a bottom view of the lower clamshell component of the first clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10E may be configured to receive one or more scalpels, one or more syringes, one or more needles, and one or more vials. In some embodiments, the recesses of the clamshell component shown in FIG. 10E may be configured to contain two or more sizes of tools, such as a small, medium, or large scalpel, a small, medium, or large syringe, a small, medium, or large needle, and a small, medium, or large vial. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10E.

[0159] FIG. 10F is a top view of the upper clamshell component of a second clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10F may be configured to receive one or more nozzles, one or more syringes, and one or more needles. In some embodiments, the recesses of the clamshell component shown in FIG. 10F may be configured to accommodate two or more sizes of tools, such as small, medium, or large nozzles, small, medium, or large syringes, and small, medium, or large needles. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10F.

[0160] FIG. 10G is a bottom view of the upper clamshell component of a second clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10G may be configured to receive one or more nozzles, one or more syringes, and one or more needles. In some embodiments, the recesses of the clamshell component shown in FIG. 10G may be configured to accommodate two or more sizes of tools, such as small, medium, or large nozzles, small, medium, or large syringes, and small, medium, or large needles. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10G.

[0161] FIG. 10H is a top view of the lower clamshell component of a second clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10H may be configured to receive one or more nozzles, one or more syringes, and one or more needles. In some embodiments, the recesses of the clamshell component shown in FIG. 10H may be configured to accommodate two or more sizes of tools, such as small, medium, or large nozzles, small, medium, or large syringes, and small, medium, or large needles. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10H.

[0162] FIG. 10I is a bottom view of the lower clamshell component of a second clamshell tool set pair. In some embodiments, the recesses of the clamshell component shown in FIG. 10I may be configured to receive one or more nozzles, one or more syringes, and one or more needles. In some embodiments, the recesses of the clamshell component shown in FIG. 10I may be configured to accommodate two or more sizes of tools, such as small, medium, or large nozzles, small, medium, or large syringes, and small, medium, or large needles. In some embodiments, the vial may contain an enzyme solution or a buffer solution. Other embodiments may have more or fewer recesses configured to receive tools or may include recesses configured to receive different tools than those shown in FIG. 10I.

[0163] 10J is a top view of the lower clamshell component of the first clamshell tool set pair shown containing tools, in the embodiment shown in FIG. 10J, a viewer can grasp tools such as one or more scalpels, one or more syringes, one or more needles, and one or more vials.

[0164] 10K is a top view of the lower clamshell component of a second clamshell tool set pair shown containing tools. In the embodiment shown in FIG. 10K, a viewer can grasp tools such as one or more nozzles, one or more syringes, and one or more needles.

[0165] Figure 11 is a top, front, right-side perspective view of the base unit, showing the base unit, housing, tissue processing area, housing base, front cover, heating element, docking spindle, display, two menu buttons, side cartridge alignment lugs, rear cartridge alignment lugs, base front plate, housing top, and control feedback sensors.

[0166] FIG. 12 is a front view of the base unit of FIG.

[0167] FIG. 13 is a rear view of the base unit of FIG.

[0168] 14 is a left side view of the base unit of FIG. 11. FIG.

[0169] 15 is a right side view of the base unit of FIG. 11. FIG.

[0170] FIG. 16 is a top view of the base unit of FIG.

[0171] FIG. 17 is a bottom view of the base unit of FIG.

[0172] Figure 18 is a top, front, right-side perspective view of the cartridge. Figure 11 shows the cartridge assembly, cartridge top cover, raised processing area, upper processing shelf, four different openings, and two cartridge tabs.

[0173] FIG. 19 is a front view of the cartridge of FIG.

[0174] FIG. 20 is a rear view of the cartridge of FIG.

[0175] FIG. 21 is a left side view of the cartridge of FIG.

[0176] FIG. 22 is a right side view of the cartridge of FIG.

[0177] FIG. 23 is a top view of the cartridge of FIG.

[0178] FIG. 24 is a bottom view of the cartridge of FIG.

[0179] Examples and Experiments Preliminary characterization work has demonstrated the effectiveness of the present disclosure. More specifically, research results demonstrate that the present system (or "automated device" as used herein) for the automated preparation of regenerative epithelial suspensions is a novel and surprising automated system useful for preparing regenerative epithelial suspensions having properties generally comparable to those of the device disclosed in U.S. Patent No. 9,029,140 (the entire contents of which are incorporated herein by reference).

[0180] Research Background The purpose of this study was to evaluate the performance of this system for the automated preparation of regenerative epithelial suspensions by assessing device output criteria related to biological function.

[0181] The RECELL® Device is a manual system for preparing regenerative epithelial suspensions (the "Manual Device") that allows clinicians to prepare regenerative epithelial suspensions using small donor skin fragments at the point-of-care.

[0182] The manual device is a disposable, stand-alone, battery-operated device that allows medical professionals to process thin skin samples (0.006-0.008 inches) at the point of care to prepare a regenerative epithelial suspension, which can then be immediately delivered to a prepared wound bed. The regenerative epithelial suspension contains a mixed population of generally viable single cells, including keratinocytes, fibroblasts, and melanocytes, obtained by enzymatic and mechanical processing of split-thickness skin samples. The cell suspension is applied to a prepared wound bed for the treatment of acute burns and full-thickness skin defects.

[0183] The present disclosure and its automated device were designed and manufactured to overcome the challenges and drawbacks of previously known methods and devices and to simplify the preparation of regenerative epithelial suspensions.

[0184] Research Procedure Trained operators prepared regenerative epithelial suspensions using manual and automated devices. For each processing sequence, trained operators obtained two identically sized skin samples from adjacent areas of the same donor skin piece. Then, using both the manual and automated devices simultaneously, operators generated regenerative epithelial suspensions from identically sized skin samples from the same donor. The operators repeated this process a total of six times, three times using each device to create 4cm skin samples. 2 On three other occasions, each device was used to generate regenerated epithelial suspensions from 24cm skin samples. 2 A regenerated epithelial suspension was generated from each skin sample. The parameters of the regenerated epithelial suspension obtained from each device were then acquired and analyzed.

[0185] Research Considerations Overall, these analyses demonstrate that both devices successfully dispersed skin and isolated cells for both lower and upper skin sample sizes. Similar functional outputs were identified for the automated device compared to the manual device. Overall, these analyses demonstrate that both devices isolated both lower and upper skin sample sizes with comparable results.

[0186] Study Conclusions Data from this study demonstrate that the automated device produces regenerative epithelial suspensions with equivalent device output characteristics related to biological function to those prepared by the manual device for skin sample sizes at the lower and upper limits of processing.

[0187] Although certain embodiments described herein relate to tissue treatment and regenerative epithelial suspensions, the devices, systems, and methods disclosed herein are not limited to these applications, and the systems and methods disclosed herein are generally applicable to medical devices in general.

[0188] Any thresholds, limits, durations, etc. described herein are not intended to be absolute and may be approximate. Additionally, any thresholds, limits, durations, etc. described herein may be fixed or may be automatically or user-adjustable. Furthermore, as used herein, relative terms such as "exceed," "greater than," "less than," etc., with respect to a reference value are intended to include values ​​equal to the reference value. For example, exceeding a positive reference value may include being equal to or greater than the reference value. Also, as used herein, relative terms such as "exceed," "greater than," "less than," etc., with respect to a reference value are intended to include the inverse of the disclosed relationship, such as "below," "less than," "greater than," etc., with respect to the reference value. Furthermore, various process blocks may be described in terms of determining whether a value meets a particular threshold, but may equally be understood in terms of whether the value (i) is below or above the threshold, or (ii) satisfies the threshold.

[0189] It is to be understood that a feature, material, characteristic, or group described in connection with a particular aspect, embodiment, or example is also applicable to any other aspect, embodiment, or example described herein, except where incompatible. All features disclosed herein (including the accompanying claims, abstract, and drawings) or steps of any disclosed method or process may be combined in any combination, except where at least some of such features or steps are mutually exclusive. Protection is not limited to the details of any embodiment described above. Protection extends to any novel feature or step, or any novel combination of such features or steps, of the features disclosed herein (including the accompanying claims, abstract, and drawings) or steps of any disclosed method or process.

[0190] Although specific embodiments have been described, these embodiments are presented by way of example only and are not intended to limit the scope of protection. Indeed, the novel methods and systems described herein may be embodied in a variety of other forms. Furthermore, various omissions, substitutions, and modifications are possible with respect to the form of the methods and systems described herein. Those skilled in the art will recognize that, in some cases, the actual steps in the illustrated or disclosed processes may differ from those shown in the figures. In some embodiments, some of the steps described may be omitted and other steps may be added. For example, the actual steps or order of steps in the disclosed processes may differ from those shown in the figures. In some embodiments, some of the steps described may be omitted and other steps may be added. For example, various components illustrated in the figures may be implemented as software or firmware on a processor, controller, ASIC, FPGA, or dedicated hardware. Hardware components, such as a controller, processor, ASIC, FPGA, etc., may include logic circuitry. Furthermore, the features and attributes of specific embodiments of the disclosure may be combined in different ways to form additional embodiments, all of which are within the scope of the present disclosure.

[0191] While the present disclosure includes particular embodiments, examples, and applications, those skilled in the art will recognize that the present disclosure extends beyond the specifically disclosed embodiments to other alternative embodiments or applications, and to obvious modifications and equivalents of each, including embodiments that do not provide all of the features and advantages described herein. Accordingly, the scope of the present disclosure is not intended to be limited by the specific disclosure of preferred embodiments herein, but may be defined by the claims as presented herein or in the future.

[0192] Conditional language such as "can" or "could," "might" or "may," unless specifically stated otherwise or understood otherwise within the context of use, conveys a general intent that certain embodiments include certain features, elements, or steps while other embodiments do not. Thus, such conditional language does not imply a general intent that any feature, element, or step is required for one or more embodiments, regardless of whether it is included in or performed in any particular embodiment, or that one or more embodiments necessarily include decision logic with or without user input or user direction. The terms "comprising," "including," "having," and the like, are synonymous and used inclusively in an open-ended manner and do not exclude additional elements, features, acts, operations, etc. Additionally, the term "or" is used in its inclusive (rather than exclusive) sense, so that, for example, when used in conjunction with elements of a list, it means one, some, or all of the elements in the list. Furthermore, as used herein, the term "each" may refer to any subset of the set of elements to which the term "each" applies, in addition to its ordinary meaning.

[0193] Unless specifically stated otherwise, conjunctions such as the phrase "at least one of X, Y, and Z" are otherwise understood as conveying that, by the context of common usage, an item, term, etc., can be X, Y, or Z. Thus, such conjunctions do not generally imply that a particular embodiment requires the presence of at least one of X, at least one of Y, and at least one of Z.

[0194] As used herein, expressions of degree such as "approximately," "about," "generally," and "substantially" refer to a value, amount, or characteristic that is close to the stated value, amount, or characteristic and still performs a desired function or achieves a desired result. For example, the terms "approximately," "about," "generally," and "substantially" can refer to an amount that is within 10%, within 5%, within 1%, within 0.1%, and within 0.01% of the stated amount.

[0195] The scope of the present disclosure is not intended to be limited by the specific disclosure of preferred embodiments in this section or elsewhere herein, but may be defined by the claims as presented in this section or elsewhere herein or in the future. The language of the claims is to be interpreted broadly based on the language employed in the claims and is not intended to be limited to the examples set forth herein or during the prosecution process, which should be construed as non-exclusive.

Claims

1. 1. A system for the automated preparation of a regenerative epithelial suspension, comprising: a base unit comprising a tissue processing area; a cartridge configured to be received in the tissue processing area, a cover including an opening configured to receive a tissue sample; a well plate located below the cover and configured to rotate relative to the cover; a cartridge comprising: a tissue dissociation device configured to mechanically dissociate tissue when the tissue sample is placed in the cartridge; A system comprising:

2. The system of claim 1 , further comprising a heating element disposed within the base unit.

3. The system of claim 2 , wherein the heating element is configured to generate sufficient heat to increase enzymatic efficiency of an enzyme disposed within the well plate.

4. The system of claim 1 , wherein the tissue dispersion device comprises a pestle.

5. The system of claim 1 , wherein the base unit comprises a tissue dispersion shaft.

6. The system of claim 5 , wherein the dispersion shaft is configured to be attached to the tissue dispersion device.

7. The system of claim 1 , further comprising a cup having a screen disposed within the cup.

8. The system of claim 7 , wherein the screen is oriented laterally within the cup.

9. The system of claim 7 or 8, wherein the opening comprises a raised portion and is configured to receive the cup.

10. 10. The system of claim 1, wherein the well plate comprises wells and is configured to rotate to align the wells with the openings.

11. 12. The system of claim 10 or 11, wherein the well plate comprises a plurality of wells and is configured to rotate to align each of the wells with the opening.

12. 12. The system of claim 11, wherein the well plate comprises a buffer well configured to receive a buffer and an enzyme well configured to receive an enzyme.

13. 13. The system of claim 12, wherein the well plate is configured to rotate to position the enzyme well below the opening when an enzyme is used to dissociate the tissue.

14. 14. The system of claim 12 or 13, wherein the well plate is configured to rotate to position the buffer wells below the openings when buffer is applied to the tissue.

15. 15. The system of any one of claims 1 to 14, further comprising a processor configured to operate the tissue disperser and rotate the well plate.

16. 1. A system for the automated preparation of a regenerative epithelial suspension, comprising: a base unit comprising a tissue processing area; a cartridge configured to be received in the tissue processing area, a cover including a raised treated opening configured to receive a cup; a well plate located below the cover and configured to rotate relative to the cover, the well plate having wells; a cartridge comprising: a tissue dissociation device configured to mechanically dissociate tissue when a tissue sample is placed in the cup; A system comprising:

17. 17. The system of claim 16, wherein the well plate has at least a first well configured to receive an enzyme solution and a second well configured to receive a buffer solution, and is configured to rotate to align the first well or the second well with the raised treatment opening.

18. 18. The system of claim 16 or 17, wherein the cup is configured to be lowered and raised into the wells of the well plate.

19. 19. The system of any one of claims 16 to 18, wherein the base unit comprises a heating element disposed within a housing.

20. 20. The system of any one of claims 16 to 19, wherein the cartridge comprises the cup and a screen disposed within the cup.

21. A cartridge for the preparation of a regenerative epithelial suspension, comprising: a cover having a raised opening; a cup configured to be received within the raised treatment opening, the cup comprising a screen; a well plate disposed beneath the cover, the well plate having wells and configured to align the wells with the raised processing openings upon rotation relative to the cover; A cartridge comprising:

22. 22. The cartridge of claim 21, wherein the screen is disposed laterally within the cup.

23. 23. A cartridge according to claim 21 or 22, wherein the screen is configured to separate particulates greater than 100 microns in size.

24. 24. A cartridge according to any one of claims 21 to 23, wherein the screen is configured to separate particulates greater than 50 microns in size.

25. 25. A cartridge according to any one of claims 21 to 24, wherein the screen is configured to separate particulates greater than 10 microns in size.

26. 26. The cartridge of any one of claims 21 to 25, further comprising a docking spindle configured to connect to a base unit.

27. 27. The cartridge of claim 26, wherein the docking spindle is configured to rotate the well plate.

28. 28. The cartridge of any one of claims 21 to 27, wherein the well comprises raised treatment elements configured to enhance dispersion of the tissue sample.

29. 30. The cartridge of claim 28, wherein the raised processing elements are configured to enhance dispersion of a tissue sample subjected to normal, rotational, or abrasive forces.

30. 30. A cartridge according to any one of claims 21 to 29, wherein the cup is configured to be positioned within the well by being raised or lowered within the raised treatment opening.

31. 31. The cartridge of any one of claims 21 to 30, wherein the cover comprises a plurality of openings.

32. 32. The cartridge of claim 31, wherein the cover comprises a buffer opening.

33. 33. The cartridge of any one of claims 21 to 32, wherein the well plate comprises a plurality of openings.

34. 34. The cartridge of claim 33, wherein the well plate comprises a buffer well and an enzyme well.

35. 35. The cartridge of claim 34, wherein the well plate is configured to be rotated to position the buffer well directly beneath the buffer opening.

36. 36. A cartridge according to claim 34 or 35, wherein the well plate is configured to be rotated to position the enzyme well directly beneath the raised treatment opening.

37. 37. The cartridge of any one of claims 21 to 36, wherein the cup is configured to receive a tissue dispersion device.

38. A cartridge for the preparation of a regenerative epithelial suspension, comprising: a cover including an opening configured to receive a tissue sample; a well plate disposed beneath the cover, the well plate having wells and configured to align the wells with the openings upon rotation relative to the cover; A cartridge comprising:

39. 39. The cartridge of claim 38, wherein the cartridge is configured to lower the tissue sample into the well aligned with the opening.

40. 40. A cartridge according to claim 38 or 39, wherein the well plate comprises buffer wells and enzyme wells and is configured to rotate to align the buffer wells or the enzyme wells with the openings.

41. 41. A cartridge according to any one of claims 38 to 40, further comprising a cup receivable in the opening in the cover and movable between a raised position above the well and a lowered position within the well.

42. 1. A method for the automated preparation of a regenerative epithelial suspension, comprising: The processor Cover and a cup containing a tissue sample; a well plate located directly under the cover, the well plate comprising a first well containing a quantity of enzyme solution and a second well containing a quantity of buffer solution; receiving an initiation signal indicating that a cartridge comprising: operating a tissue disperser on the tissue sample in the presence of the enzyme solution while the cup is positioned in the first well; raising the cup to an upper position; rotating the well plate to position the second well directly below the cup; lowering the cup to a lower position within the second well; operating the tissue dispersion device on the tissue sample in the presence of the buffer solution while the cup is positioned in the second well; A method comprising:

43. 43. The method of claim 42, wherein the tissue disperser exerts a rotational force on the tissue sample.

44. 44. The method of claim 42 or 43, wherein the tissue disperser exerts an abrasive force on the tissue sample.

45. 45. The method of any one of claims 42 to 44, wherein the tissue dispersion device comprises a pestle.

46. 46. ​​The method of any one of claims 42 to 45, wherein the tissue dispersion device comprises raised surface elements.

47. 47. The method of any one of claims 42 to 46, wherein the tissue dispersion device comprises a plurality of protrusions.

48. 48. The method of any one of claims 42 to 47, wherein the tissue sample is disposed along a screen disposed within the cup.

49. 49. The method of claim 48, wherein the screen is disposed laterally within the cup.

50. 50. The method of claim 48 or 49, wherein the screen is configured to separate particulates greater than 100 microns in size.

51. 50. The method of claim 48 or 49, wherein the screen is configured to separate particulates greater than 50 microns in size.

52. 50. The method of claim 48 or 49, wherein the screen is configured to separate particulates greater than 10 microns in size.

53. 53. The method of any one of claims 42 to 52, further comprising starting a housing motor configured to operate the tissue dispersion device.

54. 54. The method of claim 53, wherein the housing motor is configured to raise and lower the cup.

55. 55. The method of any one of claims 42 to 54, further comprising pressing the tissue dispersion device against the tissue sample multiple times when the cups are placed in the first well and the second well.

56. 56. The method of any one of claims 42 to 55, further comprising actuating the tissue dispersion device on the tissue sample when the cup is in the upper position.

57. 57. The method of any one of claims 42 to 56, further comprising starting a base plate motor configured to rotate the well plate.

58. 58. The method of any one of claims 42 to 57, further comprising rotating the well plate to bring the cup into fluid contact with a second amount of buffer solution.

59. 59. The method of claim 58, wherein the second amount of buffer solution is contained in a third well of the well plate.

60. 60. The method of claim 59, further comprising operating the tissue dispersion device on the tissue sample in the presence of the second amount of buffer solution while the cup is disposed in the third well, wherein the third well comprises one or more raised treatment elements.

61. 61. The method of any one of claims 42 to 60, further comprising receiving a completion signal indicating the presence of a regenerative epithelial suspension.

62. 62. The method of any one of claims 42 to 61, wherein the tissue dissociation device operates on the tissue sample in the presence of the enzyme solution with the cup positioned in the first well for a time effective to at least partially dissociate the tissue sample.

63. 63. The method of any one of claims 42 to 62, wherein the tissue dissociation device operates on the tissue sample in the presence of the enzyme solution with the cup positioned in the first well for a time effective to at least partially dissociate the tissue sample.

64. 64. The method of any one of claims 42 to 63, wherein the tissue dissociation device is acted on the tissue sample in the presence of the buffer solution with the cup disposed in the second well with an amount of force effective to at least partially dissociate the tissue sample.

65. 65. A non-transitory tangible computer readable medium having stored thereon computer executable instructions which, when executed by a computer processor, cause the medium to perform the method of any one of claims 42 to 64.

66. 1. A method of treating a tissue site, comprising: collecting a tissue sample containing keratinocytes and placing it in a cartridge comprising a well plate; placing the cartridge in a base unit comprising a tissue dispersion device; activating the base unit to rotate the well plate and operate the tissue disperser, wherein the well plate rotates to align one or more wells in the well plate with the tissue sample and the tissue disperser, and operation of the tissue disperser dissociates the tissue sample to form a regenerative epithelial suspension; delivering the regenerative epithelial suspension to a tissue site to enhance healing of the tissue site; A method comprising:

67. 67. The method of claim 66, wherein the tissue sample comprises a skin sample.

68. 68. The method of claim 66 or 67, wherein the regenerative epithelial suspension comprises a mixed population of viable cells.

69. 69. The method of any one of claims 66 to 68, wherein the regenerative epithelial suspension comprises fibroblasts.

70. 70. The method of any one of claims 66 to 69, wherein the regenerative epithelial suspension comprises melanocytes.

71. 71. The method of any one of claims 66 to 70, wherein the tissue site is a burned tissue site.

72. 72. The method of any one of claims 66 to 71, wherein the tissue site is a full thickness skin defect.

73. 73. The method of any one of claims 66 to 72, wherein the cartridge further comprises a cover comprising a raised treatment opening.

74. 74. The method of claim 73, further comprising placing the tissue sample in a cup disposed within the raised processing opening.

75. 75. The method of claim 74, wherein actuation of the base unit causes the cup to be lowered into a well of the well plate.

76. 76. The method of any one of claims 66 to 75, wherein the tissue dispersion device is a pestle.

77. 77. The method of any one of claims 66 to 76, wherein the well plate comprises an enzyme well.

78. 78. The method of claim 77, further comprising disposing an enzyme in the enzyme well, the enzyme being configured to dissociate tissue.

79. 79. The method of claim 77 or 78, wherein the well plate is positioned such that the enzyme well is positioned directly beneath the tissue sample.

80. 80. The method of any one of claims 77 to 79, wherein activation of the base unit causes the tissue aggregator to act on the tissue sample in the presence of an enzyme in the enzyme well.

81. 81. The method of any one of claims 66 to 80, wherein the well plate comprises a buffer well.

82. 82. The method of claim 81, further comprising disposing a buffer in the buffer well.

83. 83. The method of claim 81 or 82, wherein actuation of the base unit causes rotation of the well plate to position the buffer well directly beneath the tissue sample.

84. 84. The method of claim 83, wherein activation of the base unit causes the tissue aggregator to act on the tissue sample in the presence of the buffer in the buffer well.

85. 85. The method of any one of claims 66 to 84, wherein the well comprises one or more raised treatment elements.

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