Apparatus and system for preparing meat products

Abstract

To provide a method for forming cell-based meat and a method for growing a cell-based meat product.SOLUTION: A method for forming cell-based meat includes: providing an enclosure 310 having a cavity 316 and substrates 330 in such a way that at least one part of the substrates is non-horizontal and non-perpendicular relative to a ground surface; and flowing cells suspended in fluid media over the substrate so that at least one part of the cells adheres to the substrates to form the cell-based meat.SELECTED DRAWING: Figure 3A

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of U.S. Provisional Application No. 62 / 938,087, filed November 20, 2019, the contents of which are incorporated herein by reference in their entirety. [Background technology]

[0002] Devices constructed to support the growth and differentiation of cells in culture are typically designed to recover products of the cells secreted into the medium (e.g., antibodies), while the cells themselves are simply a by-product that is not recovered. Furthermore, these conventional devices are not designed to support the growth of continuous cell sheets (e.g., cell-based meat products) at a practical scale suitable for consumption. Thus, systems, devices, and methods for preparing meat products are desired. Summary of the Invention

[0003] Described herein are devices and systems for preparing cell-based meat products, as well as methods of using the same. Generally, the devices, systems, and methods described herein grow meat products in one or more enclosed substrates. For example, an apparatus for preparing a meat product may include a housing including a cavity, and a substrate disposed within the cavity, the substrate including a plurality of nested surfaces curved about a longitudinal axis, the surface configured to support the growth of the meat product. The substrate may be configured to receive a fluid substantially parallel to the longitudinal axis.

[0004] In some embodiments, the housing can be configured to be placed on the ground such that the longitudinal axis is at an acute angle with respect to the ground. In some embodiments, the acute angle can be at least about 85 degrees. In some embodiments, the separator can be configured to separate the meat product from the substrate in the form of a continuous sheet of cells held together by one or more cell-cell interactions and extracellular matrix produced by the cells. In some embodiments, the separator can include one or more fluid nozzles configured to spray fluid toward the substrate at a predetermined angle. In some embodiments, the separator can be configured to rotate about a predetermined axis when the fluid is sprayed onto the meat product. In some embodiments, the fluid can be sprayed at a linear velocity between about 0.003 m / s and about 3.0 m / s.

[0005] Also described herein is an apparatus for preparing a meat product that can include a housing defining a cavity, a substrate disposed within the cavity and configured to support growth of the meat product, and a fluid diffuser coupled to the housing and configured to distribute a fluid to the substrate in a predetermined flow pattern. A ratio of a diameter of the fluid diffuser to a diameter of the substrate can be between about 3:4 and about 1:1.

[0006] In some embodiments, the fluid diffuser can be configured to allow for a substantially uniform or laminar flow across a surface of the substrate. The fluid diffuser can include one or more openings. The cavity can include a headspace, and the fluid diffuser can be disposed within the headspace of the housing.

[0007] In some embodiments, the fluid diffuser may comprise a tapered surface configured to distribute the fluid to the substrate in a predetermined flow pattern. In some embodiments, the tapered surface comprises one or more of a concave shape, a conical shape, a frustum shape, a step shape, and a flare shape. In some embodiments, the tapered surface may be angled at up to about 80 degrees relative to the substrate.

[0008] In some embodiments, the fluid diffuser can include a bicone including a first side and a second side opposite the first side, The first side can be configured to receive the fluid and the second side can be configured to receive the fluid from the first side and distribute the fluid to the substrate.

[0009] Also described herein are methods of preparing an edible meat product, which may include growing the edible meat product on a substrate of the device, and separating the meat product from the substrate in the form of a continuous sheet of cells held together by one or more cell-to-cell interactions and an extracellular matrix produced by the cells.

[0010] In some embodiments, separating the edible meat products includes spraying the fluid at a predetermined angle relative to the spiralled substrate. In some embodiments, the predetermined angle can be between about 0 degrees and about 10 degrees relative to the substrate. In some embodiments, the spiralled substrate can be disposed within a cavity of the apparatus. In some embodiments, the fluid can be removed from the apparatus prior to separating the edible meat products from the substrate. In some embodiments, the substrate can have a spiral shape.

[0011] In some embodiments, an apparatus for preparing a meat product can include a housing defining a cavity and a fluid diffuser coupled to the housing. The fluid diffuser has a diameter of at least about 700 μm. 2 The cavity may include a plurality of openings having a total area of ​​about 100 mm. A plurality of substrates are configured for growing the meat products. The substrates may be disposed within the cavity and coupled to the fluid diffuser.

[0012] In some embodiments, the fluid diffuser may include at least a first portion and a second portion. The first portion may be angled between about 90 degrees and about 170 degrees relative to the second portion. In some embodiments, the first portion and the second portion are each angled at up to about 80 degrees relative to the plurality of substrates.

[0013] In some embodiments, the plurality of openings of the fluid diffuser are substantially equally spaced. In some embodiments, each of the plurality of openings has a diameter of at least about 30 μm. In some embodiments, the fluid diffuser includes an interface between a first portion and a second portion, the interface including one or more of the plurality of openings. In some embodiments, the fluid diffuser is coupled to a proximal end of a plurality of substrates.

[0014] In some embodiments, the fluid diffuser comprises one or more of stainless steel, ceramic, and polymer. In some embodiments, the first portion and the second portion are substantially flat. In some embodiments, the housing and the fluid diffuser are substantially circular and each have an inner diameter and an outer diameter. The outer diameter of the fluid diffuser is substantially the same as the inner diameter of the housing.

[0015] In some embodiments, the fluid diffuser is configured to output a substantially uniform, laminar flow of fluid to a plurality of substrates. In some embodiments, the fluid diffuser is configured to releasably engage the housing. In some embodiments, the fluid diffuser is disposed at least partially within the cavity.

[0016] In some embodiments, the device has a plurality of fluid channels. In some embodiments, at least one of the fluid channels is associated with at least one of the plurality of substrates. In some embodiments, the one or more fluid channels are planar. In some embodiments, each of the fluid channels is between adjacent substrates. In some embodiments, fluid is configured to flow unidirectionally through the plurality of fluid channels. In some embodiments, the plurality of fluid channels are substantially parallel to one another. In some embodiments, the fluid channels include a space between adjacent substrates of between about 0.3 mm and about 5.0 cm.

[0017] In some embodiments, the substrates are parallel to one another. In some embodiments, one or more of the substrates are planar. In some embodiments, one or more of the substrates are non-planar. In some embodiments, the substrates include one or more of stainless steel, ceramic, and polymer. In some embodiments, the apparatus includes up to about 10,000,000 substrates. In some embodiments, the substrates include one or more of the following shapes: flat, rectangular, rod, ball, and disk. In some embodiments, the width of each substrate is between about 10 cm and about 400 cm. In some embodiments, each substrate is about 430 cm. 2 From about 8,000cm 2 In some embodiments, the plurality of substrates are configured to releasably engage the housing. In some embodiments, the one or more substrates are configured to grow meat products on both sides of the substrate.

[0018] In some embodiments, at least one collector is coupled to the housing. In some embodiments, the collector is at least about 25 μm 2 In some embodiments, the collector is coupled to a distal end of the plurality of substrates. In some embodiments, the collector comprises one or more of stainless steel, ceramic, polymer. In some embodiments, the collector is configured to receive meat products separated from the plurality of substrates. In some embodiments, the collector is configured to releasably engage the housing.

[0019] In some embodiments, the housing comprises one or more inlets configured to receive a fluid. In some embodiments, the housing comprises one or more outlets configured to output a fluid. In some embodiments, the one or more inlets are disposed on a first side of the housing and the one or more outlets are disposed on a second side of the housing opposite the first side of the housing. In some embodiments, the housing has a volume between about 25 L and about 20,000 L.

[0020] In some embodiments, the one or more outlets have a diameter configured to output meat products separated from the plurality of substrates. In some embodiments, the housing has a substantially cylindrical shape. In some embodiments, a rotor is coupled to the housing and configured to rotate the housing about a transverse axis of the housing.

[0021] Also described herein is a system for preparing a meat product, including an apparatus including a plurality of substrates, a fluid pump coupled to the apparatus, a fluid source coupled to the fluid pump, and a controller coupled to the fluid pump, the controller configured to generate a first fluid pump signal to grow the meat products on the substrates and generate a second fluid pump signal to separate the meat products from the substrates.

[0022] In some embodiments, the first fluid pump signal is configured to provide a substantially uniform, laminar flow of fluid to a plurality of substrates using the fluid pump, in some embodiments, the first fluid pump signal indicates a first flow rate and the second fluid pump signal indicates a second flow rate that is greater than the first flow rate.

[0023] In some embodiments, the one or more sensors are configured to measure one or more parameters of the fluid. In some embodiments, the first fluid pump signal indicates a flow rate of up to about 3.0 meters per second. In some embodiments, the second fluid pump signal corresponds to a linear velocity in the device of up to about 10 meters per second. In some embodiments, the fluid source includes a growth medium. In some embodiments, the growth medium includes cells. In some embodiments, the system is configured to grow meat on the substrate for at least about 1 day.

[0024] Also described herein is a method of preparing a meat product that includes providing a plurality of non-human cells to at least one substrate of the device, culturing the cells on the substrate using a first fluid comprising a growth medium to produce a meat product, and separating the meat product from the substrate using the first fluid or a second fluid flowing through the device.

[0025] In some embodiments, culturing the cells comprises providing a substantially uniform and layered first fluid to the substrate. In some embodiments, the substrate comprises about 5,000 cells / cm. 2 and approximately 100,000 cells / cm 2 is supplied to the cells.

[0026] In some embodiments, the first fluid is recirculated through the device at a velocity between about 0.0001 meters per second and about 3.0 meters per second. In some embodiments, the first fluid flows through the device from a first height to a second height that is less than the first height. In some embodiments, the cells are cultured on the substrate for at least one day. In some embodiments, the first fluid configures a planar flow through the substrate. In some embodiments, the meat product is held together by extracellular matrix secreted by the cells and / or cell-cell interactions.

[0027] In some embodiments, the cells include one or more of myoblasts, mesangioblasts, myofibroblasts, mesenchymal stem cells, hepatocytes, fibroblasts, pericytes, adipocytes, epithelium, chondrocytes, osteoblasts, osteoclasts, pluripotent cells, somatic stem cells, and endothelial cells.

[0028] In some embodiments, the cells include one or more cells from livestock, poultry, game, and aquatic animal species. In some embodiments, when culturing the cells, the substrate is aligned substantially vertically to the ground. In some embodiments, the apparatus includes rotating the substrate so that the substrate is parallel to the ground to deliver the one or more cells to the substrate.

[0029] In some embodiments, the separated meat products are received in a collector. In some embodiments, the collector is disposed within a cavity of the apparatus. In some embodiments, the collector is coupled to the exterior of the apparatus. In some embodiments, the substrate meat products are within about 0.0001 mm. 3 About 0.1m from 3In some embodiments, the substrate is sterilized after separating the meat product. In some embodiments, after sterilizing the substrate, another meat product is cultured and separated. In some embodiments, sterilizing the substrate comprises steaming the apparatus. In some embodiments, at least one of the substrates is removed from the apparatus.

[0030] Also described herein are methods of preparing a meat product that include any of the devices and systems described herein. In some embodiments, the fluid diffuser is removable from the device. In some embodiments, the collector is removable from the device. Also described herein are meat products produced by the methods described herein.

[0031] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication and color drawing(s) will be provided by the Office upon request and payment of the necessary fee. This application can be understood by reference to the following description in conjunction with the accompanying drawings. [Brief description of the drawings]

[0032] [Figure 1] FIG. 1 is a block diagram illustrating an exemplary embodiment of a system for preparing a meat product. [Figure 2A] FIG. 2A is an exemplary perspective view of an exemplary embodiment of an apparatus for preparing a meat product. [Figure 2B] FIG. 2B is an exemplary side view of the device shown in FIG. 2A. [Figure 2C] FIG. 2C is another exemplary embodiment of an apparatus for preparing a meat product. [Figure 2D] FIG. 2D is an exemplary side view of the device shown in FIG. 2C. [Figure 2E] FIG. 2E is another exemplary side view of the device shown in FIG. 2C. [Figure 3A] FIG. 3A is an exemplary cross-sectional side view of an exemplary embodiment of an apparatus for preparing a meat product. [Figure 3B] FIG. 3B is an exemplary perspective view of the internal components of the device shown in FIG. 3A. [Figure 3C] FIG. 3C is an exemplary cross-sectional perspective view of the internal components of the device shown in FIG. 3B. [Figure 3D] FIG. 3D is an exemplary side view of the internal components of the device shown in FIG. 3B. [Figure 4A] FIG. 4A is an exemplary perspective view of an exemplary embodiment of a fluid diffuser. [Figure 4B] FIG. 4B is an exemplary top view of the fluid diffuser shown in FIG. 4A. [Figure 4C] FIG. 4C is another exemplary perspective view of the fluid diffuser shown in FIG. 4A. [Figure 4D] FIG. 4D is an exemplary side view of the fluid diffuser shown in FIG. 4A. [Figure 5A] FIG. 5A is an exemplary perspective view of an exemplary embodiment of a plurality of substrates. [Figure 5B] FIG. 5B is an exemplary side view of the substrate shown in FIG. 5A. [Figure 6A] FIG. 6A is an exemplary perspective view of an exemplary embodiment of a collector. [Figure 6B] FIG. 6B is an exemplary plan view of the collector shown in FIG. 6A. [Figure 6C] FIG. 6C is an exemplary side view of the collector shown in FIG. 6A. [Figure 7A] , [Figure 7B] , [Figure 7C] , [Figure 7D] 7A, 7B, 7C, and 7D are exemplary perspective views of an exemplary embodiment of an assembly process of an apparatus for preparing meat products. [Figure 8] 8A, 8B, and 8C are exemplary side views of an exemplary embodiment of a rotational process for an apparatus for preparing meat products. [Figure 9A] , [Figure 9B]9A and 9B are exemplary cross-sectional side views of fluid flow through an exemplary embodiment of an apparatus for preparing meat products. [Figure 10A] FIG. 10A is an exemplary perspective view of an exemplary embodiment of an apparatus for preparing a meat product. [Figure 10B] FIG. 10B is an exemplary front view of the device shown in FIG. 10A. [Figure 10C] FIG. 10C is an exemplary side view of the internal components of the device shown in FIG. 10B. [Figure 11A] FIG. 11A is an exemplary perspective view of an exemplary embodiment of a fluid diffuser. [Figure 11B] FIG. 11B is an exemplary side view of the fluid diffuser shown in FIG. 11A. [Figure 11C] FIG. 11C is an exemplary top view of the fluid diffuser shown in FIG. 11A. [Figure 12A] FIG. 12A is an exemplary perspective view of an exemplary embodiment of a plurality of substrates. [Figure 12B] FIG. 12B is an exemplary side view of the substrate shown in FIG. 12A. [Figure 12C] FIG. 12C is an exemplary front view of the substrate shown in FIG. 12A. [Figure 12D] FIG. 12D is an exemplary plan view of the substrate shown in FIG. 12A. [Figure 13A] FIG. 13A is an exemplary cross-sectional side view of an exemplary embodiment of an apparatus for preparing a meat product. [Figure 13B] FIG. 13B is another exemplary side view of the device shown in FIG. 13A. [Figure 13C] FIG. 13C is an exemplary perspective view of the device shown in FIG. 13A. [Figure 13D] FIG. 13D is an exemplary cross-sectional perspective view of the device shown in FIG. 13A. [Figure 13E] FIG. 13E is another exemplary cross-sectional perspective view of the device shown in FIG. 13A. [Figure 13F] FIG. 13F is an exemplary plan view of the device shown in FIG. 13A. [Figure 14A] FIG. 14A is an exemplary top view of an exemplary embodiment of a fluid diffuser. [Figure 14B] FIG. 14B is an exemplary perspective view of the fluid diffuser shown in FIG. 14A. [Figure 14C] FIG. 14C is an exemplary side view of an exemplary embodiment of a fluid diffuser and holder. [Figure 14D] FIG. 14D is an exemplary perspective view of the fluid diffuser and holder shown in FIG. 14C. [Figure 15A] , [Figure 15B] , [Figure 15C] , [Figure 15D] 15A, 15B, 15C, and 15D are exemplary perspective views of exemplary embodiments of fluid diffusers. [Figure 16A] FIG. 16A is an exemplary perspective view of an exemplary embodiment of a substrate. [Figure 16B] FIG. 16B is an exemplary side view of the substrate shown in FIG. 16A. [Figure 16C] FIG. 16C is an exemplary plan view of the substrate shown in FIG. 16A. [Figure 16D] FIG. 16D is an exemplary detailed plan view of the substrate shown in FIG. 16C. [Figure 16E] FIG. 16E is an exemplary top view of the substrate shown in FIG. 16A disposed within a housing. [Figure 16F] FIG. 16F is an exemplary schematic diagram of an exemplary embodiment of a substrate. [Figure 16G] FIG. 16G is an exemplary schematic diagram of another exemplary embodiment of a substrate. [Figure 17A] FIG. 17A is an exemplary schematic diagram of an exemplary embodiment of a separator of an apparatus for preparing a meat product. [Figure 17B] , [Figure 17C] 17B and 17C are exemplary schematic diagrams of exemplary embodiments of separator and substrate configurations. [Figure 18A] , [Figure 18B] , [Figure 18C] , [Figure 18D] , [Figure 18E] 18A, 18B, 18C, 18D, and 18E are exemplary cross-sectional side views of fluid flow through an exemplary embodiment of an apparatus for preparing meat products. [Figure 19] FIG. 19 is an exemplary flow chart of an exemplary method for preparing a meat product using an apparatus of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0033] Detailed Description The systems, devices, and methods provided herein are useful for controlling the growth, attachment, detachment, and / or retention properties of cultured cells (e.g., cell sheets) for preparing meat products. In general, the systems, devices, and methods described herein grow and attach large numbers of cells to one or more substrates for a predetermined period of time to produce a meat product. In some embodiments, the number and / or size of substrates can be adjusted to produce a predetermined amount of meat product (e.g., as part of a commercial-scale process to form a cell-based meat product, also referred to herein as edible tissue).

[0034] Certain challenges faced in large-scale cell growth for the production of edible meat are addressed by the systems, devices, and methods provided herein. Conventional devices, such as cell culture vessels and bioreactors, are utilized to cultivate cells in suspension culture and are not suitable for meat production because they are unable to sustainably grow cell sheets. Other conventional devices designed for the growth of adherent cells are limited in size and incorporate tortuous fluid channels to accommodate those size constraints. In contrast, the devices described herein can be cost-effectively sized to grow cells (e.g., cell sheets) of sufficient physical dimensions to generate meat products. For example, in some embodiments, the growth of one or more cell sheets in the devices of the present disclosure requires sufficient time for the cells to remain attached to the substrate for a predetermined period of time (e.g., growth period) and / or for the cell state (e.g., observed metabolic signatures are changes in cell differentiation). One or more fluids (e.g., growth medium, culture medium, liquid) can flow through the device in a predetermined flow pattern to promote cell growth / differentiation / attachment on one or more substrates. However, it is difficult to provide a predetermined fluid flow over a large surface area. As described herein, the fluid diffuser can be configured to receive a fluid and distribute the fluid in a substantially uniform, laminar flow to one or more substrates to promote consistent cell growth across the surface area of ​​the one or more substrates.

[0035] Once the cells have grown into cell sheets on one or more substrates (e.g., production of meat products), it is desirable to recover the grown meat products from the device, preferably at a predetermined size, substantially intact, and / or contemporaneous, for further processing. In some embodiments, recovery of the one or more cell sheets is assisted by a fluid release mechanism (e.g., fluid-based shear stress), thereby enabling controlled and large-scale production and collection of edible meat products. In some embodiments, a fluid other than the cell growth application can be used to separate the generated cell sheets from the substrate as an end product for collection. For example, the separator can be configured to receive and distribute another fluid to the one or more substrates in a predetermined flow pattern sufficient to separate the grown meat products from the substrate. Thus, the devices described herein provide significant improvements in one or more of cell sheet growth, attachment, retention, and separation in the device compared to conventional systems and techniques. Additionally, the device can be configured to be disassembled, sterilized, and reused, reducing waste and costs and improving production efficiency.

[0036] As used herein, a cell sheet can be either a single layer of cells or multiple layers of cells. A cell sheet can include an extracellular matrix (ECM). In some embodiments, a cell sheet can be held together by ECM produced by the cells, and / or the cells can form a sheet via cell-cell junction proteins to form a substantially continuous unit. For example, secretion of sufficient ECM proteins allows the constituent cells to attach to each other as a substantially continuous cell sheet through a combination of cell-cell and cell-ECM interactions. In some embodiments, the cell sheet is held together by an endogenously produced scaffold, such as an ECM. In other embodiments, the ECM is provided exogenously.

[0037] In some embodiments, a method of preparing a meat product may include providing a plurality of non-human cells to a substrate of an apparatus, culturing the cells on the substrate using a first fluid comprising a growth medium to produce a meat product (e.g., a cell sheet), and separating the meat product from the substrate using a second fluid flowing through the apparatus (e.g., through one or more separators).

[0038] I. Equipment Provided herein is an apparatus for controlling the growth of a cell sheet (meat product) on an encapsulated substrate and / or the detachment of the meat product from the substrate.

[0039] An apparatus of the present disclosure for preparing a meat product includes (a) a housing (e.g., chamber, housing, vessel) defining a cavity, (b) a fluid diffuser coupled to the housing having a plurality of openings, and (c) a plurality of substrates configured to grow meat products, the plurality of substrates being disposed within the cavity and coupled to the fluid diffuser. In some embodiments, the apparatus may further include a separator configured to separate the meat products from the one or more substrates, and a collector coupled to the housing for collecting the meat products.

[0040] The components of the device (e.g., housing, fluid diffuser, substrate, separator, collector) may be composed of materials including, but not limited to, one or more of polychlorotrifluoroethylene, polyetherimide, polysulfone, polystyrene, polycarbonate, polypropylene, silicone, polyetheretherketone, polymethylmethacrylate, nylon, acrylic, polyvinyl chloride, vinyl, phenolic resins, petroleum-derived polymers, glass, polyethylene, terephthalate, stainless steel, titanium, aluminum, cobalt chromium, chromium, silicates, glasses, alloys, ceramics, carbohydrate polymers, mineral-like substances, and combinations or composites thereof.

[0041] These components are described in further detail in turn herein. Non-limiting exemplary embodiments of the apparatus are illustrated and described in more detail with reference to Figures 1, 3A-3D, 7A-7D, 8A-8C, 10A-10C, and 17A-17C. Figures 2A-9B and 17A-17C show a first exemplary embodiment of an apparatus for preparing a meat product. Figures 2A-2E, 10A-12B, and 17A-17C illustrate a second exemplary embodiment of an apparatus for preparing a meat product. Figures 2A-2E and 13-17C show a third exemplary embodiment of an apparatus for preparing a meat product.

[0042] A. Chassis In general, the housing of the device may be configured to provide a sealed chamber to allow for aseptic growth of the meat product. The housing may include one or more inlets configured to receive a fluid and one or more outlets configured to output the fluid. In some embodiments, the one or more inlets may be located on a first side of the housing and the one or more outlets may be located on a second side of the housing opposite the first side of the housing.

[0043] In some embodiments, the device can have dimensions that include an internal cavity volume of at least about 1 L. For example, the device can have an internal volume of between about 25 L and about 200 L, between about 100 L and about 500 L, between about 500 L and about 1,000 L, between about 1,000 L and about 40,000 L, including all values ​​and subranges therebetween. In some embodiments, the cavity can include a head space. The fluid diffuser can be disposed within the head space of the housing.

[0044] Non-limiting exemplary embodiments of the housing are illustrated and described in more detail with reference to Figures 2A-2E, 3, 7A-7D, 8A-8C, 9A, 13A-13C, 13E-13F, and 18A-18E.

[0045] B. Fluid Diffuser The apparatus of the present disclosure includes one or more fluid diffusers (also referred to herein as flow conditioners or flow regulators) configured to receive a fluid and provide a predetermined fluid flow to one or more substrates. For example, the fluid diffuser can be configured to receive and distribute the fluid to a plurality of substrates in a predetermined fluid flow pattern, such as a substantially uniform laminar flow, to promote consistent cell growth across the surface of the one or more substrates. For example, the fluid diffuser can be configured to receive the fluid from an inlet and provide a linear fluid flow to the one or more substrates. In this manner, the fluid can be introduced into the cavity of the housing through a relatively narrow inlet port and distributed evenly across one or more substrates having relatively wide dimensions (e.g., diameter, length, width).

[0046] In some embodiments, the flow of fluid through the fluid diffuser may be gravity assisted. In some embodiments, the device may include an inlet configured to receive the fluid and one or more outlets configured to output the fluid. Thus, in such embodiments, the fluid diffuser may be coupled between the inlet and one or more substrates. In some embodiments, the fluid diffuser may be a separate component from the inlet or may be integrated with the inlet. The fluid diffuser may be disposed within the headspace of the housing.

[0047] In some embodiments, the fluid diffuser can have a dimension (e.g., diameter) that is smaller than a dimension of the substrate. In some embodiments, the fluid diffuser can include a ratio of a dimension of the fluid diffuser to a dimension of the substrate (e.g., diameter, length, width) that is between about 3:4 and about 1:1.

[0048] In some embodiments, the fluid diffuser may include a tapered surface configured to distribute the fluid to the substrate in a predetermined flow pattern (e.g., to distribute the fluid approximately evenly across the diameter of the substrate). The tapered surface may include one or more of a concave, conical, frustum, stepped, and flared shape. For example, the tapered surface may be angled at up to about 80 degrees relative to the substrate.

[0049] In some embodiments, the fluid diffuser may include a bicone. For example, the bicone may include a first side and a second side opposite the first side. The first side may be configured to receive a fluid and the second side may be configured to receive the fluid from the first side and distribute the fluid to the substrate. The fluid diffuser may be configured to allow for a substantially uniform, or laminar, flow across a diameter of the substrate.

[0050] In some embodiments, the fluid diffuser can be radially symmetric. In some embodiments, the longitudinal axis of the fluid diffuser can be substantially perpendicular to the substrate. In some embodiments, the fluid diffuser and the substrate have a predetermined distance of up to about 20 cm. For example, the fluid diffuser can be positioned higher (e.g., above) relative to the substrate such that the fluid flows downward from the fluid diffuser to the substrate. In the case of a substrate having a spiral (e.g., scroll) configuration, the fluid can flow in parallel along the height of the substrate, passing through each turn of the spiral.

[0051] In some embodiments, the fluid diffuser may include a plurality of openings. In some embodiments, the openings are at least about 700 μm 2 The apertures may include a total area of ​​about 100 μm. The apertures may be any shape (e.g., circular, polygonal, slits, and combinations thereof). In some embodiments, the apertures are approximately circular in shape, with each aperture having a diameter of about 30 μm, allowing the growth medium to pass through the fluid diffuser. In other embodiments, the diameter of the circular apertures may include between about 30 μm and about 10 cm, between about 0.5 cm and about 3 cm, and all values ​​and subranges therebetween. In some embodiments, the multiple apertures of the fluid diffuser may be substantially equally spaced. In some embodiments, the size and / or frequency of the apertures may vary as a function of distance from the center of the fluid diffuser.

[0052] In some embodiments, the fluid diffuser may be comprised of only one portion. In other embodiments, the fluid diffuser may have two or more portions (e.g., at least a first portion and a section portion). In some embodiments, one or more portions of the fluid diffuser may be substantially flat. In other embodiments, the first and second portions are adjacent to each other, and these adjacent portions of the fluid diffuser are angled with respect to each other, for example, between about 90 degrees and about 170 degrees, including all values ​​and subranges therebetween. For example, the greater the angle between the portions, the lower the height of the fluid diffuser may be. Furthermore, the aforementioned portions of the fluid diffuser may be angled up to about 80 degrees with respect to the substrates. In some embodiments, the fluid diffuser has an interface between adjacent portions, for example, the interface that constitutes the third portion. The interface may selectively have an opening.

[0053] In some embodiments, the fluid diffuser can be configured to releasably engage with the housing and can be disposed at least partially within the cavity, for example, the fluid diffuser can be disposed within a headspace of the housing (e.g., the open space above the substrate).

[0054] In some embodiments, the device may have multiple fluid diffusers. In some embodiments, the fluid diffusers may be positioned above and / or below the substrate. For example, a fluid diffuser positioned above the substrate may distribute fluid downward to the substrate by gravity, while a fluid diffuser positioned below the substrate may distribute fluid upward to the substrate against gravity.

[0055] Non-limiting exemplary embodiments of fluid diffusers are illustrated in and described in more detail with reference to Figures 1, 3A-3D, 4A-4C, 7A-7B, 9A, 10A-10C, 11A-11C, 13A-13F, 14A-14D, 15A-15D, and 18A-18E.

[0056] C. Base material The multiple substrates in the apparatus of the present disclosure allow for meat products to be grown at high density. For example, the substrates may include a surface configured to promote cell attachment, differentiation, and / or growth to form edible meat products. Once grown to a predetermined size, the grown meat products may be separated from each substrate as described in more detail herein. The substrates of the present disclosure may be of any predetermined size.

[0057] In some embodiments, the substrate disposed within the cavity may be rolled about an axis (e.g., the longitudinal axis of the housing or substrate) to allow for denser tissue growth and increase the structural rigidity of the substrate than a flat substrate. The increased rigidity of the substrate due to the curvature allows the substrate to be thinned accordingly, which may further increase the volume available to support the growth of meat products. For example, the substrate may be disposed within the cavity and may include multiple nested surfaces (e.g., spiral-shaped, scroll-like substrates) that are curved about a longitudinal axis and have surfaces configured to support the growth of meat products. For example, the nested surfaces may include one tube within at least one other tube (e.g., inner tube, outer tube). The scroll-like substrate may be rotated about a longitudinal axis. In some embodiments, the scroll-like substrate may have an outer diameter of at least about 1 cm. In some embodiments, the substrate may have a thickness of at least about 1 μm.

[0058] In some embodiments, the scrolled substrate can be wound more than once around a given axis. For example, the substrate can have multiple revolutions about the axis. That is, the substrate can be wound multiple times around itself to form a general spiral shape. Each revolution of the spiral can have a configuration in which the substrate goes completely around itself (e.g., relative to the innermost edge of the substrate). In some embodiments, the substrate can be bent more than 360 degrees around the axis (e.g., to wrap or form more than two revolutions). In some embodiments, the substrate can be bent around the longitudinal axis of the housing or substrate.

[0059] In some embodiments, adjacent perimeters (e.g., layers, surfaces) of a substrate may include a predetermined spacing configured to aid in fluid flow and meat product growth. For example, the space between each perimeter of the substrate may allow for fluid flow and meat product growth on the surface of the substrate. That is, fluid channels may be formed between adjacent perimeters.

[0060] In some embodiments, the substrate may include a first substrate portion and a second substrate portion formed separately from the first substrate portion. An end of the first substrate portion may be proximate to an end of the second substrate portion. In some embodiments, an end of the first substrate portion may be bonded to an end of the second substrate portion (e.g., bonded in an end-to-end configuration). In some embodiments, an end of the first substrate portion may overlap an end of the second substrate portion. In some of these embodiments, the substrate may include three or more substrate portions bonded in any combination.

[0061] In some embodiments, a scroll-like substrate (e.g., a spiral-like substrate) can be configured to be self-spacing in that the peripheries have a predetermined spacing relative to one another. For example, the substrate can be formed with a predetermined curvature such that it naturally forms a spiral shape. One or more protrusions on the surface of the substrate can ensure that each periphery of the substrate is generally separated by a predetermined spacing. In some embodiments, the surface of the substrate can have one or more multiple protrusions having a height of at least about 0.5 mm. Adjacent (e.g., adjacent) protrusions can include a predetermined spacing of up to about 20 cm along the surface of the substrate. The predetermined spacing can be based on the radius of the substrate. For example, the predetermined spacing can increase from the innermost portion of the substrate to the outermost portion, or vice versa. The one or more protrusions can have a linear or nonlinear shape. In some embodiments, the one or more protrusions can be imprinted on the surface of the substrate.

[0062] In some embodiments, the substrate may be angled (e.g., inclined) relative to the ground (e.g., the ground). The angled substrate may promote growth of meat products on a surface of the substrate compared to a substrate positioned perpendicular to the ground. For example, a surface of the substrate facing away from the ground may be configured to aid in adhesion, differentiation, and growth of meat products. In some embodiments, the substrate and housing may be angled relative to the ground. In some embodiments, the housing may be configured to rest on the ground. The substrate may define a longitudinal axis that forms an acute angle with the ground. In some embodiments, the acute angle may be at least about 85 degrees and may include 90 degrees.

[0063] In some embodiments, each substrate may have a dimension between about 10 cm and about 400 cm, including all values ​​and subranges therebetween, hi some embodiments, the plurality of substrates may include up to about 10,000,000 substrates, including all values ​​and subranges therebetween.

[0064] In some embodiments, the spacing between adjacent (eg, adjacent) substrates can be between about 0.3 mm and about 5.0 cm, and all values ​​and subranges therebetween.

[0065] In some embodiments, each substrate has a surface area of ​​up to about 150,000,000 cm 2 , and all values ​​and subranges therebetween.

[0066] The substrates described herein may be sized and shaped to be placed in and / or integrated with the device. In some embodiments, multiple substrates may be used in a given arrangement. In other embodiments, multiple substrates may be arranged in a parallel plate configuration (e.g., adjacent to each other) that allows for a multi-layered cell sheet to be formed within the device. For example, the parallel plate configuration allows for perfusion of fluid (e.g., growth medium, nutrients) between the plates. In some embodiments, the geometry of the fluid channels (described further below) may include spaces between adjacent substrates.

[0067] In some embodiments, the substrate may be substantially planar. Additionally or alternatively, one or more of the substrates may be non-planar. For example, each substrate may have one or more of the following shapes: rectangle, rod, ball, disk, spiral, coil, helix, corrugated, and sinusoidal shapes (e.g., Raschig ring super ring). For example, a plurality of spherical balls configured to fill a cavity of the housing may have both high density and large surface area for the growth of meat products. The corresponding spaces between the spherical balls constitute the fluid channels of the device. The substrates need not be parallel to each other, but may be angled or intersecting each other. The surface of the substrate may include one or more channels, grooves, and depressions configured as fluid channels. In some embodiments, the substrate may be extended in the direction of one or more dimensions (e.g., length, width, diameter) of the housing to better utilize the interior volume of the housing.

[0068] In some embodiments, the substrates may be arranged in an ordered or unordered arrangement. For example, multiple rod substrates may be arranged in a random pattern. Multiple substrates may include multiple shapes (e.g., plates and rods, balls and coils). One or more substrates may be arranged in a symmetrical or asymmetrical arrangement.

[0069] In some embodiments, the substrate may be configured to grow meat products on both sides of the substrate, which may increase the amount of meat product that is grown on the substrate.

[0070] The substrates described herein support the growth and maintenance of cells, including, but not limited to, cells of one or more of endoderm, mesoderm, ectoderm, and combinations thereof. In some embodiments, the cells include one or more cells from livestock (e.g., cattle, pigs, sheep, goats), poultry (e.g., birds), game animals, aquatic species, and the like. In some embodiments, the cells include one or more of myoblasts, mesangioblasts, myofibroblasts, mesenchymal stem cells, hepatocytes, fibroblasts, pericytes, adipocytes, epithelium, chondrocytes, osteoblasts, osteoclasts, pericytes, pluripotent stem cells, somatic stem cells, and endothelial cells. The cell types described herein further encompass any of their differentiation states. For example, the cells include myoblasts, myotubes, mature skeletal muscle, fibroblasts, tissues containing cells and secreted extracellular matrix, adipocytes, adipose tissue, epithelial cells, epithelial tissue, vascular endothelium, combinations thereof, and the like. In some embodiments, the cells may include vertebrate cells or non-vertebrate cells. In some embodiments, the cells may include non-mammalian cells (e.g., insect cells, avian cells, fish cells, reptile cells, invertebrate cells). In some embodiments, the cells may be genetically modified from their original state (e.g., gene insertion, deletion, or recombination). Examples of genetic modifications include cells engineered to overexpress myogenic transcription factors. The cells may be present, for example, in different ratios. In some embodiments, the devices, systems, and methods disclosed herein include those described in International Publication No. WO 2015 / 066377, filed October 30, 2014, International Publication No. WO 2017 / 124100, filed January 17, 2017, International Publication No. WO 2018 / 208628, filed May 5, 2018, International Publication No. WO 2019 / 014652, filed July 13, 2018, International Application No. PCT / US2020 / 034949, filed May 28, 2020, and U.S. Patent Application No. 62 / 938,087, filed November 20, 2019, the contents of each of which are incorporated herein by reference in their entirety.

[0071] In other embodiments, the substrate can be disposed within and / or in combination with an exogenously provided three-dimensional lattice-like configuration (e.g., a scaffold) (e.g., comprising a three-dimensional, porous and / or lattice-like structure).

[0072] In other embodiments, the substrate does not have an exogenous scaffold and is scaffold-free (e.g., excluding three-dimensional porous and / or lattice-like structures that are not endogenously produced by the cells). Thus, in such embodiments, the cell sheet can be grown in the absence of an exogenous, non-naturally secreted scaffold structure (but can be grown in the presence of, e.g., naturally secreted, endogenously produced extracellular matrix (ECM)).

[0073] The substrates described herein may be comprised of one or more of a solid material and a semi-solid material (eg, a hydrogel).

[0074] In some embodiments, the substrate may include a textured surface to promote cell / cell sheet adhesion, differentiation, and growth. In some embodiments, one or more substrates may be engineered to enhance one or more properties (e.g., coated to improve adhesion).

[0075] Non-limiting exemplary embodiments of the substrate are shown in and described in more detail with reference to Figures 1, 3A-3D, 5A-5B, 7A-7D, 8A-8C, 9A-9B, 10A-10C, 12A-12D, 13A-13F, 16A-16F, and 18A-18E.

[0076] D. Fluid Channel In general, the disclosed devices have a plurality of fluid channels. At least one of the fluid channels is associated with at least one of the plurality of substrates. A liquid may be configured to flow unidirectionally through the plurality of fluid channels. For example, the fluid flows from a higher position to a lower position along a predetermined path (e.g., a linear path). In some embodiments, the fluid channels may be formed between adjacent substrates or adjacent circumferences (turns, layers, surfaces, etc.) of a scroll-like substrate. For example, a spiral substrate may constitute a spiral fluid channel. In some embodiments, the substrate may have one or more spacing features (e.g., protrusions) configured to maintain a predetermined width of the fluid channel. The plurality of fluid channels may have the same or different shapes and / or dimensions.

[0077] In some embodiments, each fluid channel can have a width from about 0.3 mm to about 5.0 cm, and all values ​​and subranges therebetween.

[0078] In some embodiments, one or more of the fluid channels can be planar. In some embodiments, each fluid channel can be between adjacent substrates. In some embodiments, the multiple fluid channels can be approximately parallel to one another.

[0079] Non-limiting exemplary embodiments of fluid channels are shown in and described in more detail with reference to Figures 1, 3A-3D, 5A-5B, 7A-7D, 8A-8C, 9A-9B, 10A-10C, 12A-12D, 13A-13F, 16A-16F, and 18A-18E.

[0080] E. separator In some embodiments, the device may include one or more separators configured to separate the meat products from the plurality of substrates in a predetermined manner. For example, a separator, such as a fluid nozzle, may be configured to receive and distribute a fluid (e.g., liquid, water) to one or more substrates in a predetermined flow pattern to separate the grown meat products from the substrates, such that the meat products may be collected in portions having a predetermined size. For example, the meat products may be separated and collected from the device as a continuous tissue sheet (e.g., tissue sheet maintains integrity) and maintains a desired shape or texture, rather than a collection of disjointed cells. For example, the continuous tissue sheet may be slid off the substrate for collection. In some embodiments, sterility of the system may be maintained while separating the tissue from the substrate, thereby increasing efficiency and decreasing run time. For example, the separator may be configured to operate without opening the housing.

[0081] In some embodiments, the one or more separators may be configured integral with or separate from the housing of the device. For example, a first separator may be disposed within a sidewall of the housing and a second separator may be disposed within a cavity of the housing. For example, the second separator may be coupled (e.g., fixed) to a fluid diffuser, a substrate, or other components disposed within the device. The one or more separators may be configured to translate and / or rotate relative to the substrate to aid in the meat separation process. For example, the separator may be configured to move radially between the center and periphery of the housing to distribute fluid to a larger volume of substrate. Additionally or alternatively, the separator may be configured to rotate to distribute fluid in a predetermined (e.g., cyclical) pattern to separate meat from a larger volume of substrate.

[0082] FIG. 17A is a schematic diagram of an apparatus (1700) including a plurality of separators (1740). The apparatus (1700) may include a housing (1710) including an inlet (1712) and an outlet (1714) and may further define a cavity (1716). A fluid diffuser (1720) and at least one substrate (1730) may be disposed within the cavity (1716). In some embodiments, one or more separators (1750) (e.g., fluid nozzles) may be coupled to an inner wall of the housing (1710). For example, the separators (1750) may be integral with the housing (1710) and protrude from the housing (1710) into the cavity (1716). The separators (1750) may be configured to distribute fluid to the substrate (1730) in a predetermined fluid flow pattern (1750). In some embodiments, the fluid flow pattern may include one or more of a fan, spray, jet, column, solid stream, full cone, hollow cone, mist, intermittent, droplet, rotation, or static pattern. For example, the fluid flow pattern may include a small diameter with a relatively high pressure (e.g., focused stream) or a large diameter with a relatively low pressure (e.g., diffuse spray). In some variations, the fluid flow may have a pressure between about 0 PSI and about 50 PSI. For illustration, FIG. 17A shows a separator (1740) with a fluid flow pattern (1750) having a fan pattern. For example, the fluid (1750) may be injected parallel and / or perpendicular to the interface between the meat product and the substrate (1730), as described in more detail with reference to FIGS. 17B and 17C. In some embodiments, the separator (1750) may be disposed in one or more of the sidewalls, top, bottom, lid, headspace (e.g., the empty space above the substrate (1730)), etc. of the housing (1710).

[0083] The fluid diffuser (1720) may be disposed between the inlet (1712) and at least one substrate (1730). For example, the fluid diffuser (1720) may be disposed within a headspace of the housing (1710). Any of the fluid diffuser (1720), substrate (1730), and separator (1740) may be removable or fixed to the housing (1710). In some embodiments, one or more fluids (e.g., liquid, growth medium, gas) may be configured to flow from the inlet (1712) toward the outlet (1714). The fluid diffuser (1720) may be configured to receive and distribute the fluid to one or more substrates (1730) in a predetermined flow pattern. In some embodiments, the housing (1710) may include multiple inlets, outlets, and separators (1740).

[0084] Fluid dispensed from the separators (1750) can be configured to flow in any predetermined direction and / or pattern. For example, a first separator can be configured to dispense fluid from a higher position to a lower position (e.g., downward) to an innermost portion of the substrate (1730) (e.g., the center of FIG. 17A), and a second separator can be configured to dispense fluid from a side (e.g., a sidewall of the housing (1710)) to an outermost portion of the substrate (1730).

[0085] 17B and 17C are schematic perspective views of separator configurations relative to a substrate. In particular, FIG. 17B shows a separator (1742) (e.g., a fluid nozzle) configured to distribute a fan-shaped fluid flow pattern (1752) perpendicular to the substrate (1730) (e.g., across the thickness of the substrate (1730)). For example, the perpendicular flow pattern (1752) forms a flat fan-shaped spray pattern (e.g., a fluid sheet) that intersects with each of the parallel plates (or volute perimeters) of the substrate (1730). In some embodiments, the separator can be configured to spray fluid onto the substrate at a predetermined angle of up to about 10 degrees relative to the substrate. For example, a portion of the substrate (1730) directly below the separator (1742) can receive the fluid at 0 degrees, and another portion of the substrate (1730) can receive the fluid at a predetermined angle. In some embodiments, the separator (1742) can be set against and / or moved across the substrate (1730) to distribute the spray pattern across a larger surface area of ​​the substrate (1730). The separator (1742) in a vertical configuration shown in FIG. 17B can be configured to separate tissue on both sides of the substrate. For example, tissue that does not directly receive the fluid stream from the nozzle can be separated from the substrate (1730) by the jet of the fluid stream.

[0086] FIG. 17C illustrates a separator (1744) (e.g., a fluid nozzle) configured to distribute a fan-shaped fluid flow pattern (1754) parallel to the substrate (1730). For example, the parallel flow pattern (1754) forms a flat fan-shaped spray pattern (e.g., a fluid sheet) that intersects with one of the parallel plates (or circumferences) of the substrate (1730) (e.g., the rearmost plate in FIG. 17C). In FIG. 17C, the portion of the substrate (1730) directly below the separator (1744) receives the fluid at 0 degrees. In some embodiments, the separator (1744) can be wrapped and / or positioned perpendicular to the entire substrate (1730) to distribute a spray pattern across each plate (or circumference) of the substrate (1730).

[0087] In some embodiments, the fluid flow pattern may have an angle of up to about 120 degrees. For example, the fluid may configure a sector-shaped fluid flow having an angle of up to about 120 degrees. In some embodiments, to reduce the velocity of the fluid flow, the substrate is not submerged (e.g., not flooded) in the fluid when receiving the fluid flow having the predetermined fluid flow pattern. That is, at least a portion of the fluid may be drained from the housing prior to tissue separation. In some embodiments, the substrate may receive the fluid in a column (e.g., having a circular cross section) parallel to the substrate. In some embodiments, the fluid may have a linear velocity of between about 0.003 m / s and about 0.3 m / s, between about 0.03 m / s and about 0.3 m / s, between about 0.03 m / s and about 3.0 m / s, between about 0.3 m / s and about 3.0 m / s, including all values ​​and subranges therebetween. In some embodiments, the separator may be configured to rotate (e.g., pitch, yaw, roll) about a predetermined axis as the fluid is sprayed onto the meat product and substrate. In some embodiments, the separator may be a distance away from the substrate of up to about 10 cm, up to about 15 cm, up to about 30 cm, up to about 50 cm, including all values ​​and subranges therebetween. In some embodiments, the tissue may be separated from the substrate via indirect contact with the fluid stream. For example, the redirected fluid stream from the sidewall of the housing may have sufficient energy to separate the tissue from the substrate.

[0088] Non-limiting exemplary embodiments of the separator are shown and described in more detail in Figures 17A-17C. Any of the system and device embodiments described herein can include one or more separators.

[0089] F. collector In some embodiments, the apparatus may further include one or more collectors coupled to the housing of the apparatus to receive and collect the meat products from the plurality of substrates.

[0090] In some embodiments, the collector of the present disclosure has a plurality of openings, e.g., at least about 25 μm 2For example, each aperture may have a diameter between about 30 μm and about 2.5 cm, including all values ​​and subranges therebetween.

[0091] In some embodiments, the collector may be a separate component from the housing or may be integrally configured with the housing. In some embodiments, the collector may be configured to releasably engage the housing and may be bonded to an end remote from the plurality of substrates. In some embodiments where the collector is external to the housing, the collector may be at least about 20 μm 2 The aperture includes one or more openings having an area of

[0092] Non-limiting exemplary embodiments of the collector are illustrated and described in more detail with reference to Figures 1, 3A-3D and 6A-6C.

[0093] II. system Systems are described that include one or more components necessary to produce meat products using the devices described herein. For example, the systems described herein may support, grow, separate, and harvest one or more cell sheets cultured on the substrate of the device. In general, the systems described herein include one or more of the devices of the present disclosure, fluid pumps, fluid sources, and controllers (including memory, processors, and computer instructions).

[0094] In some embodiments, an apparatus for preparing a meat product may include a housing including a cavity and a substrate disposed within the cavity. The substrate may include a convolution and a surface configured to aid in the growth of the meat product. A fluid diffuser may be coupled to the housing and configured to distribute a fluid to the substrate in a predetermined flow pattern. In some embodiments, a ratio of dimensions of the fluid diffuser to dimensions of the substrate may be between about 3:4 and about 1:1.

[0095] In some embodiments, an apparatus for preparing a meat product may include a housing defining a cavity and a substrate disposed within the cavity. The substrate may be configured to support growth of the meat product. A fluid diffuser including a tapered surface may be configured to distribute a fluid to the substrate in a predetermined flow pattern.

[0096] In general, a system for preparing a meat product includes: (a) an apparatus including one or more substrates, (b) a fluid pump coupled to the apparatus, (c) a fluid source coupled to the fluid pump, and (d) a controller coupled to the fluid pump, the controller configured to generate a first fluid pump signal to generate a meat product in the substrate and a second fluid pump signal to separate the meat product from the substrate. For example, the fluid pump is configured to provide a fluid flow of a growth medium (e.g., culture medium) to the apparatus. The controller is configured to control the flow rate and other conditions (e.g., temperature, pressure) of the fluid within the apparatus.

[0097] In some embodiments, the first fluid pump signal can be configured to provide a substantially uniform and laminar flow of fluid to the plurality of substrates using the fluid pump. In an exemplary embodiment, the first fluid pump signal includes a first flow rate and the second fluid pump signal includes a second flow rate that is higher than the first flow rate. In some of these embodiments, the first fluid pump signal includes a flow rate of up to about 3.0 meters per second and the second fluid pump signal corresponds to a linear velocity in the device of up to about 10 meters per second. The fluid source typically includes a growth medium and / or cells. The system is configured to grow the meat on the substrate for at least about one day, but may extend to three days, five days or more depending on the nature of the cells being grown. For example, the meat may be grown for one week or more. Optionally, one or more sensors of the system are configured to measure one or more parameters of the condition of the fluid and the cell sheet.

[0098] 1 is an exemplary block diagram of an embodiment of a system (100) including a device (110), a fluid pump (130), a fluid source (140), a controller (150), and a sensor (160). In some embodiments, the device (110) includes one or more of an inlet (112), a fluid diffuser (114), a substrate (116), cells (118), a collector (120), an outlet (122), a rotor (124), and a separator (126). One or more fluid pumps (130) may be in fluid communication with the device (110) and the fluid source (140). The fluid source (140) (e.g., a fluid reservoir) is configured to store a fluid (142) (e.g., growth medium, cells, liquid, and combinations thereof). The fluid pump (130) is configured to pump and / or recirculate a fluid (142) through the fluid conduits (132) that form a closed loop path of the system (100). The system (100) may include one or more fluid sources (140) and fluids (142). For example, a growth medium may be used to grow meat products within the device (110), while a separation fluid (e.g., water) may be used to separate the grown meat products from the substrate (116).

[0099] A. fluid As used herein, the fluids that may be circulated throughout the system may include one or more of growth media (e.g., cell culture media), nutrients, metabolites, signaling factors, liquids, water, gases, and compositions configured for growing meat products. In some embodiments, the fluids may further include cells that are used to seed the substrate on which the edible meat products are grown. In some embodiments, the fluids may include additives including one or more of amino acids, antifoam agents, shear protectants, and proteins.

[0100] In some embodiments, the cell culture medium may include liquid cell culture medium or dehydrated cell culture medium. Dehydrated cell culture medium or dehydrated powder medium (DPM) may include a mixture of amino acids, salts, glucose, and other chemicals necessary for cell growth. DPM may be hydrated with a liquid, such as purified water, to form a liquid cell culture medium.

[0101] In some embodiments, the cell culture medium may include a complete composition sufficient to grow cells by itself without other additives. That is, the complete cell culture medium includes all components necessary for the growth of cells grown in a bioreactor. In some embodiments, the complete cell culture medium may include one or more of water, buffer, and nutrients that aid in the growth of cells. In some embodiments, the cell culture medium may include an incomplete composition configured to grow cells by adding at least one other composition. For example, an incomplete cell culture medium composition may be formed by omitting a component that is sensitive to oxidation. In some variations, the pH of the incomplete cell culture medium may be different from the pH of the complete cell culture medium.

[0102] B. Fluid Pump

[0103] In some embodiments, one or more fluid pumps are coupled to a fluid conduit in fluid communication with the apparatus to generate a predetermined fluid flow rate through the apparatus to aid in at least one of growing the meat products and separating the meat products from the substrate. In some embodiments, the fluid pumps may include one or more of a positive displacement pump (e.g., a peristaltic pump), a centrifugal pump, combinations thereof, etc. One or more fluid sources may be coupled to the fluid pumps.

[0104] In some embodiments, the fluid pump can be configured to output a first fluid flow, which may be regulated by a fluid diffuser to provide a substantially uniform and laminar fluid flow to the plurality of substrates, and a second fluid flow configured to separate the meat products from the substrates. In some embodiments, the second fluid flow may have a flow rate of up to about 10 meters per second. The fluid pump may be configured to operate for an extended period of time (e.g., days, weeks). For example, the fluid pump can be configured to pump fluid for at least one day (e.g., 3, 5, 10, 15, 20, 25, 30 days).

[0105] C. fluid source In some embodiments, one or more fluid sources (e.g., fluid reservoir, gas generator) can be coupled to one or more fluid pumps and devices for preparing a meat product. The fluid source can be configured to store a fluid. The fluid source can include a growth medium containing one or more cells. For example, the fluid source can be configured to store different fluid compositions for different growth stages of the meat product. The fluid source can be configured to store a recirculated medium and separately store fresh medium. In some embodiments, one or more sensors can be coupled to the fluid source to measure one or more parameters of the fluid, such as pH, dissolved gas concentration, osmolality, turbidity, hydration, conductivity, absorbance, nutrient concentration, waste concentration, ion concentration, oxygen concentration, temperature, etc.

[0106] In some embodiments, the fluid source may be configured to retain air bubbles generated by the fluid pump to prevent air bubbles from forming in the fluid conduit. That is, the air bubbles may be retained in the fluid source upstream of the conduit. For example, the fluid source may include a cavity configured to retain a volume of fluid such that the fluid fills the cavity to a predetermined height that is higher than the height of the inlet and outlet of the fluid source. The space above the predetermined height in the cavity of the fluid source may contain air. During use, the operation of the pump may generate one or more air bubbles that may enter the fluid source. When air bubbles enter the fluid source, the air within the air bubbles may be retained within the fluid source such that bubble-free fluid may be output from the outlet of the fluid source to the device. This may prevent air bubbles from forming in the conduit of the device and may promote uniform and laminar fluid flow through the device.

[0107] D. Control Unit In general, the systems described herein may include at least one apparatus for preparing a meat product and a corresponding controller coupled to a fluid pump and a sensor. In some embodiments, the sensor may be configured to generate signal data. The signal data may be received by the controller and used to generate a fluid pump signal for controlling the fluid pump.

[0108] As shown in FIG. 1 , in some embodiments, the controller (150) may be coupled to one or more of the device (110), the fluid pump (130), the fluid source (140), and the sensor (160). The controller (150) may comprise one or more of a processor (152), a memory (154), an input device (156), and a communication device (158). In some embodiments, the controller (150) may be configured to receive data from one or more of the device (110), the fluid pump (130), the fluid source (140), and the sensor (160). In some embodiments, the sensor (160) may include one or more of a flow sensor, a temperature sensor, a pH sensor, a dissolved gas sensor, a pressure sensor, an optical sensor, a turbidity sensor, etc. In some embodiments, the data generated by the sensor (160) may be processed and used to monitor and / or control one or more components of the system (100).

[0109] Thus, the controller (150) may monitor and / or control the conditioning and / or recovery (e.g., separation) of the edible meat products. As described in more detail herein, the controller (150) may be coupled to one or more networks using a communication device (158). The controller (150) may include a processor (152) and a memory (154) coupled to an input device (156).

[0110] The controller (150) may include computer instructions running on a computer to cause the processor (152) to perform one or more operations described herein. In some embodiments, the computer instructions may be configured to cause the processor to receive signal data from the sensor, generate a fluid pump signal, and output data to a user. The controller (150) may include one or more processors (152) and one or more machine-readable memories (154) in communication with the one or more processors (152). The processor (152) may incorporate data received from the memory (154) and user inputs to control the system (100). The memory (154) may further store instructions to cause the processor (152) to execute modules, processes, and / or functions associated with the system (100). The controller (150) may connect to and control one or more of the sensors (160), the fluid pump (130), the communication device (158), etc., via wired and / or wireless communication channels.

[0111] The controller (150) may be implemented to be compatible with numerous general purpose or special purpose computing systems or configurations. Various exemplary computing systems, environments, and / or configurations that may be suitable for use with the systems and devices disclosed herein include, but are not limited to, software or other components embodied in or on servers or server computing devices, such as routing / connection components, multiprocessor systems, microprocessor-based systems, distributed computing networks, personal computing devices, network appliances, portable (e.g., handheld) or laptop devices, etc. Examples of portable computing devices include wearable computers in the form of smartphones, personal digital assistants (PDAs), mobile phones, tablet PCs, smart watches, etc., portable or wearable augmented reality devices that interface with the patient's environment via sensors and are used as head-mounted displays for visualization, eye tracking, and user input.

[0112] The processor (152) may be any suitable processing device configured to run and / or execute a sequence of instructions or code, and may include one or more of a data processor, an image processor, a graphics processing unit, a physical processing unit, a digital signal processor, and / or a central processing unit. The processor (152) may be, for example, a general purpose processor, a field programmable gate array (FPGA), an application specific integrated circuit (ASIC), combinations thereof, and the like. The processor (152) may be configured to run and / or execute application processes and / or other modules and perform processing and / or functions related to the system and / or its associated networks. The underlying device technology may be provided as a variety of component types, including metal oxide semiconductor field effect transistor (MOSFET) technologies such as complementary metal oxide semiconductor (CMOS), bipolar technologies such as emitter coupled logic (ECL), polymer technologies (e.g., silicon bonded polymer and metal bonded polymer-metal structures), mixed analog and digital, combinations thereof, and the like.

[0113] In some embodiments, the memory (154) may include a database (not shown), which may be, for example, a random access memory (RAM), a memory buffer, a hard drive, an erasable programmable read only memory (EPROM), an electrically erasable read only memory (EEPROM), a read only memory (ROM), a flash memory, combinations thereof, and the like. As used herein, a database refers to a data storage resource. The memory (154) may store instructions that cause the processor (152) to execute modules, processes, and / or functions associated with the control unit (150), such as calibration, signal processing, sensor analysis, notification, communication, authentication, user configuration, and combinations thereof. In some embodiments, the storage may be network-based and accessible to one or more authorized users. The network-based storage may be referred to as remote data storage or cloud data storage. The signal data and analysis stored in the cloud data storage (e.g., database) may be accessible to authorized users over a network such as the Internet. In some embodiments, the database may be a cloud-based FPGA.

[0114] Some embodiments described herein relate to computer storage products with non-transitory computer-readable media (also referred to as non-transitory processor-readable media) that hold instructions or computer code for performing various computer-implemented processes. The computer-readable media (or processor-readable media) is non-transitory in the sense that it does not include a transitory propagating signal itself (e.g., a propagating electromagnetic wave that carries information over a transmission medium such as space or a cable). The media and computer code (also referred to as code or algorithm) may be designed and constructed for a specific purpose.

[0115] Examples of non-transitory computer readable media include, but are not limited to, magnetic storage media such as hard disks, floppy disks, and magnetic tapes; optical storage media such as compact disks / digital video disks (CD / DVD); compact disks-read only memory (CD-ROM); holographic devices; magneto-optical storage media such as optical disks; solid-state storage media such as solid-state drives (SSD) and solid-state hybrid drives (SSHD); carrier wave signal processing modules; hardware devices specially configured to store and execute program code, such as application specific integrated circuits (ASICs), programmable logic devices (PLDs), read only memory (ROM), random access memory (RAM) devices, etc. Other embodiments described herein relate to computer program products that may include, for example, the instructions and / or computer code disclosed herein.

[0116] The systems, devices, and methods described herein may be implemented by software (executed on hardware), hardware, or a combination thereof. Hardware modules may include, for example, general purpose processors (or microprocessors or microcontrollers), field programmable gate arrays (FPGAs), application specific integrated circuits (ASICs), combinations thereof, and the like. Software modules (executed on hardware) may be expressed in a variety of software languages, including C, C++, Java, Python, Ruby, VisualBasicR, and / or other object-oriented, procedural, or other programming languages ​​and development tools. Examples of computer code include, but are not limited to, microcode or microinstructions, machine instructions such as those produced by a compiler, code used to generate web services, and files containing high level instructions executed by a computer using an interpreter. Other examples of computer code include, but are not limited to, control signals, encrypted code, and compressed code.

[0117] The input devices (156) may enable a user to directly and / or remotely interact with and / or control the system (100). For example, the input devices (156) may be configured to allow a user to input commands, and the output devices may be configured to allow the user and / or other users (e.g., technicians) to receive output related to the operation of the system (100) (e.g., displaying system data on a display device). In some embodiments, the communication devices (158) may enable the controller (150) to communicate with one or more of a network (e.g., the Internet), a remote server, and a database, as described in more detail herein.

[0118] The input device (156) may serve as a communication interface between a user (e.g., an operator) and the controller (150). In some embodiments, the input device (156) may include an output device (e.g., a touch screen and a display) and may be configured to input data from and output data from one or more of a sensor, an input device, an output device, a network, a database, and a server. For example, signal data generated by a sensor may be processed by the processor (152) and memory (154) and output visually by one or more output devices (e.g., a display). The signal data, sensor data, and / or meat product data may be received by the controller (150) and output through visual, auditory, and / or tactile feedback via one or more output devices. As another example, user control of an input device (e.g., a joystick, keyboard, touch screen) may be received by the input device (156) and then processed by the processor (152) and memory (154) to output control signals to one or more components of the system (100). In some embodiments, the input device (156) may function as both an input device and an output device (eg, a handheld controller configured to generate control signals while providing haptic feedback to a user).

[0119] The output device outputs data and may include one or more of a display device, an audio device, and a tactile device. The display device may be configured to display a graphical user interface (GUI). The input device (156) may include an integrated display and / or a video output that may be connected for output to one or more general purpose displays, including remote displays accessible via the Internet or a network. The output data may be encrypted to ensure privacy, and all or a portion of the output data may be stored on a server or database. The display device may allow a user to view signal data, calibration data, tissue data, image data, cell sample data, system data, fluid data, patient data, and / or other data processed by the controller (150). In some embodiments, the output device may include a display device including at least one of a light emitting diode (LED), a liquid crystal display (LCD), an electroluminescent display (ELD), a plasma display panel (PDP), a thin film transistor (TFT), an organic light emitting diode (OLED), an e-paper / e-ink display, a laser display, a holographic display, and combinations thereof, and the like.

[0120] The audio device may audibly output sensor data, meat product data, fluid data, pump data, system data, alerts, and / or warnings. For example, the audio device may output an audible warning upon malfunction of a fluid pump and / or a sensor. In some embodiments, the audio device may include at least one of a speaker, a piezoelectric audio device, a magnetostrictive speaker, and / or a digital speaker. In some embodiments, a user may communicate with other users using the audio device and the communication channel.

[0121] Haptic devices may be incorporated into one or more input and output devices to provide additional sensory output (e.g., force feedback) to the user. For example, a haptic device may generate a haptic response (e.g., vibration) to confirm a user input to an input device (e.g., joystick, keyboard, touch surface). In some embodiments, the haptic device may include a vibration motor configured to provide tactile feedback to the user. Additionally or alternatively, the haptic feedback may notify the user of errors such as a pump malfunction and / or a fluid interruption, thereby preventing potential harm to the system.

[0122] Some embodiments of the input device may include at least one switch configured to generate a control signal. For example, the input device may be configured to control one or more pumps to control the flow rate of a fluid. In some embodiments, the input device may include a wired and / or wireless transmitter configured to transmit the control signal to a wired and / or wireless receiver of the controller (150). For example, the input device may include a touch surface for a user to provide an input (e.g., a finger touch on the touch surface) corresponding to the control signal. An input device including a touch surface may be configured to detect touch and movement on the touch surface using any of a number of touch sensing technologies including capacitive, resistive, infrared, optical imaging, dispersive signal, acoustic pulse recognition, and surface acoustic wave technologies. In embodiments of the input device including at least one switch, the switch may include at least one of, for example, a button (e.g., hard key, soft key), a touch surface, a keyboard, an analog stick (e.g., joystick), a directional pad, a pointing device (e.g., mouse), a trackball, a jog dial, a step switch, a rocker switch, a pointer device (e.g., stylus), a motion sensor, an image sensor, and a microphone. The motion sensor can receive user movement data from the optical sensor and classify the user's gestures as control signals, and the microphone can pick up audio and recognize the user's voice as a control signal.

[0123] In some embodiments, the controller 150 may communicate with other devices via one or more wired and / or wireless networks. The communication device 156 may facilitate communication with other devices via one or more external ports (e.g., Universal Serial Bus (USB), multi-pin connectors) configured to couple directly to the other devices or indirectly via a network (e.g., the Internet, wireless LAN).

[0124] In some embodiments, the communication device (156) may include a radio frequency receiver, a transmitter, and / or an optical (e.g., infrared) receiver and transmitter configured to communicate with one or more devices and / or networks. The communication device (156) may communicate wired and / or wirelessly. In some embodiments, the communication device (156) may include a radio frequency (RF) circuit (e.g., an RF transceiver) having one or more of a receiver, a transmitter, and / or an optical (e.g., infrared) receiver and transmitter configured to communicate with one or more devices and / or networks. The RF circuit may transmit and receive RF signals (e.g., electromagnetic signals). The RF circuit converts between electrical and electromagnetic signals and communicates with communication networks and other communication devices via electromagnetic signals. The RF circuit may include one or more of an antenna system, an RF transceiver, one or more amplifiers, a tuner, one or more oscillators, a digital signal processor, a CODEC chipset, a subscriber identity module (SIM) card, a memory, etc. A wireless network may refer to any type of digital network that is not connected by any type of cable.

[0125] Examples of wireless communications in a wireless network include, but are not limited to, cellular, radio, satellite, and microwave communications. Wireless communication may use any of a number of communication standards, protocols, and technologies, including Global System for Mobile Communications (GSM), Enhanced Data GSM Environment (EDGE), High Speed ​​Downlink Packet Access (HSDPA), Wideband Code Division Multiple Access (W-CDMA), Code Division Multiple Access (CDMA), Time Division Multiple Access (TDMA), Bluetooth, Near Field Communications (NFC), Radio Frequency Identification (RFID), Wireless Fidelity (Wi-Fi) (e.g., IEEE 802.11a, IEEE 802.11b, IEEE 802.11g, IEEE 802.11n), Voice over Internet Protocol (VoIP), Wi-MAX, protocols for email (e.g., Internet Message Access Protocol (IMAP), Post Office Protocol (POP)), instant messaging (e.g., eXtensible Messaging and Presence Protocol (XMPP), Session Initiation Protocol for Instant Messaging, Presence Leveraging Extensions (SIMPLE), Instant Messaging and Presence Leveraging Extensions (SIMPLE), Instant Messaging and Presence Leveraging Extensions (SIMPLE), Instant Messaging and Presence Leveraging Extensions (TDMA), and the like. Communications protocols include, but are not limited to, Intermediate Mobile Phone System (IMPS), Short Message Service (SMS), or other suitable communications protocols. Some wireless network deployments use a combination of multiple cellular networks or a combination of cellular, Wi-Fi, and satellite communications.

[0126] In some embodiments, wireless networks may connect to wired networks to interface with the Internet, other carrier voice and data networks, business networks, and personal networks. Wired networks are typically transmitted over copper twisted pairs, coaxial cables, and / or fiber optic cables. There are various types of wired networks, including wide area networks (WANs), metropolitan area networks (MANs), local area networks (LANs), Internet area networks (IANs), campus area networks (CANs), global area networks (GANs) such as the Internet, wireless personal area networks (PANs) (e.g., Bluetooth, Bluetooth Low Energy), and virtual private networks (VPNs). As used herein, a network refers to any combination of wireless, wired, public, and private data networks that are typically interconnected via the Internet to provide an integrated networking and information access system.

[0127] E. Exemplary embodiments Exemplary embodiments of the apparatus and system are described with reference to the figures. As shown in FIG. 1, an embodiment of the apparatus (110) includes one or more of an inlet (112), a fluid diffuser (114), a substrate (116), a collector (120), an outlet (122), an optional rotor (124), and a separator (126). In some embodiments, the apparatus is rotatable about an axis of rotation (e.g., an axis of rotation parallel to the ground). The apparatus (110) can be configured for large-scale production of cell sheets forming meat products. In some embodiments, the apparatus has dimensions including an internal cavity volume of at least 1 L. For example, the apparatus (110) can have an internal volume between about 25 L and about 200 L, between about 100 L and about 500 L, between about 500 L and about 1,000 L, and between about 1,000 L and about 20,000 L, including all values ​​and subranges therebetween.

[0128] In some embodiments, the device (110) is coupled to one or more fluid pumps (130), fluid conduits (132), fluid sources (140), and sensors (160). For example, different fluid conduits (132) coupled to each fluid pump (130) and fluid reservoir (140) may be configured to circulate separate fluids (142) (e.g., different growth medium compositions, liquids, gases) to the cells (118) within the device (110) to support efficient cell growth across multiple meat product growth stages. In some embodiments, the device (110) is coupled to at least one control unit (150). For example, the optional sensors (160) may each be coupled to a sensor controller.

[0129] In some embodiments, one or more portions of the device (110), such as the housing, the fluid diffuser (114), the substrate (116), the collector (120), and the separator (126), may be designed to be reusable (e.g., used multiple times, sterilized, and reused). Additionally or alternatively, one or more components of the device (110) may be designed to be disposed of after a predetermined number of uses. In some embodiments, any of the components of the device may be substantially non-degradable.

[0130] As described herein, the devices and systems described herein can generate a variety of mechanical and hydrodynamic forces, such as interstitial flow, during long-term growth (e.g., days, weeks, months) of meat products in a confined, three-dimensional environment.

[0131] 2A-9B show an apparatus for preparing a meat product. FIGS. 2A-2E show perspective and side views, respectively, of apparatus (200, 250) for preparing a meat product. The apparatus (200) may include a housing (210) defining a cavity therein. The housing (210) may include at least one inlet (220) configured to receive fluid, for example, from a fluid conduit coupled to a fluid source (not shown), and at least one outlet (230) configured to output one or more of the fluid and a meat product (e.g., a cell sheet). For example, the outlet (230) may be of a size sufficient to output a meat product separated from one or more substrates within the housing (210). In some embodiments, the inlet (210) is disposed on a first side of the housing (210) and the outlet (230) is disposed on a second side of the housing (210) opposite the first side. In some embodiments, the at least one inlet (220) may be configured for both receiving and outputting a fluid. Similarly, the at least one outlet (230) may be configured for both receiving and outputting a fluid. For example, the flow of the fluid may begin in a first direction (e.g., from the inlet (220) to the outlet (230)) and change to the reverse direction (e.g., from the outlet (230) to the inlet (220)) during one or more steps of the method of preparing the meat product. As shown in Figures 2A-2E, the housings (210, 260) may have a generally cylindrical shape, although they are not limited by a particular shape. In some embodiments, the housings (210, 260) may have a volume of at least 25 L. For example, the housings (210, 260) may have a volume between about 100 L and about 500 L.

[0132] In some embodiments, the device (200) may include one or more inlets (220) configured to receive a fluid, such as a growth medium, a liquid, and / or a gas. For example, a fluid pump may be configured to be in fluid communication with one or more inlets (220) of the device (200) via a respective fluid conduit. In some embodiments, the housing (210) of the device (200) may define the inlets (220) along an upper surface (e.g., a top surface) of the device (200) relative to the ground. This allows fluid to flow through the device (200) from a first height (e.g., a height of the inlets (220)) to a second height (e.g., a height of the outlets (230)) that is lower than the first height. This configuration may facilitate drainage and complete circulation of the fluid through the device (200).

[0133] In some embodiments, the inlet (220) may include a valve (not shown), for example, configured to control the flow of fluid and prevent backflow of fluid into the fluid conduit. In some embodiments, the inlet (220) may be located on one or more sides (e.g., sidewalls) of the device (200). In some embodiments, the inlet (220) may be configured for different steps of the meat production process (e.g., sterilization, seeding, growth, separation). For example, different inlets may be configured to receive sterilization fluid, growth medium, and separation fluid to optimize fluid distribution throughout the vessel at different stages of the meat production process. For example, different inlets may receive growth medium with different stages (e.g., metabolically conditioned and reoxygenated states). In some embodiments, the one or more inlets (220) of the device (200) may be configured as access to one or more substrates disposed within the cavity of the housing (210). For example, the inlet (210) may be configured to receive a fluid containing cells used to seed one or more substrates. In some embodiments, the inlet (220) may comprise a connector (e.g., adapter, fitting) configured to mate with one or more fluid conduits and a fluid diffuser. In some embodiments, the outlet (230) may comprise a connector (e.g., adapter, fitting) configured to mate with one or more fluid conduits and a collector.

[0134] Similar to the device (200) shown in Figures 2A and 2B, Figures 2C-2E show a device (250) that includes a housing (260), an inlet (270), and an outlet (280). In some embodiments, the device (250) may be at an angle (e.g., off-axis) relative to an axis (290) that is perpendicular to the ground when preparing meat products. That is, the device (250) may be tilted during use. In some embodiments, the device (250) may be at an angle of up to about 5 degrees relative to the axis (290), or at least about 85 degrees relative to the ground. Growing a continuous tissue sheet at a non-perpendicular or non-parallel angle to the ground can facilitate attachment of the cell culture to the substrate while gravity aids in the flow of fluid through the substrate. This allows the size of the housing to be scaled vertically, thereby optimizing the physical footprint of the device (250). In some embodiments, the longitudinal axis of the internal components of the device (250) may be inclined at an angle relative to the axis (290), while the longitudinal axis of the housing (250) itself may be parallel to the axis (290). For example, the longitudinal axis of one or more substrates and / or fluid diffusers may be inclined relative to the axis (290), with the inlet (270) and outlet (280) each being parallel to the axis (290). In some embodiments, the housing may be configured to be placed on the ground. The longitudinal axis (290) may be at an acute angle relative to the ground. In some embodiments, the acute angle is at least about 85 degrees. In some embodiments, the surface of the substrate facing away from the ground is configured to aid in the growth of meat products. Any of the devices described herein may be used at an angle relative to an axis perpendicular to the ground.

[0135] 3A is a cross-sectional side view of an apparatus (300) for preparing a meat product. The apparatus (300) includes a housing including an inlet (312), an outlet (314), and further defining a cavity (316). Disposed within the cavity (316) are a fluid diffuser (320), a plurality of substrates (330), a collector (340), and a holder (350). In some embodiments, the holder (350) may be configured to releasably engage (e.g., retain) one or more of the fluid diffuser (320), the plurality of substrates (330), and the collector (340). The holder (350) may be coupled to an interior wall of the housing (310) via one or more supports (352). In some embodiments, the holder (350) may be integrally configured to an interior wall of the housing (310). The fluid diffuser (320) may be disposed between the inlet (312) and the plurality of substrates (330), and the collector (340) may be disposed between the plurality of substrates (330) and the outlet (314). Any of the fluid diffuser (320), substrate (330), and collector (340) may be removable or fixed to the housing (310). In some embodiments, the fluid (e.g., growth medium) may be configured to flow in a direction from the inlet (312) to the outlet (314). In some embodiments, the fluid (e.g., growth medium) may be configured to flow in a direction from the outlet (314) to the inlet (312). In some embodiments, the fluid flow may be configured to change direction through the inlet (312) and the outlet (314) during use.

[0136] FIG. 3B is a perspective view of the internal components of the device (300), excluding the housing (310) and the holder (350). FIG. 3C is a cross-sectional perspective view of the internal components of the device (300). FIG. 3D is a side view of the internal components of the device (300). The fluid diffuser (320) can be configured to distribute the received fluid flow throughout each of the substrates (330). The fluid diffuser (320) can be coupled to a proximal end of the substrates (330). In some embodiments, the fluid diffuser (320) can be configured to releasably engage the housing and at least partially disposed within the cavity (316). For example, the fluid diffuser (320) can be slidably and releasably engaged to the holder (350) without tools or fasteners, thereby aiding in the maintenance and sterilization of the device (300). The collector (340) can be configured to receive meat products and fluid from each of the substrates (330). In some embodiments, a collector (340) is coupled to a distal end of the plurality of substrates (330). Additionally or alternatively, the collector may be disposed outside of the housing (310). For example, the collector may receive the output of the outlet (314).

[0137] In some embodiments, the substrates (330) may be configured to releasably engage the housing (330). For example, the substrates (330) may slidably and releasably engage the holder (350) without tools or fasteners, thereby facilitating maintenance and sterilization of the substrates (330). In some embodiments, the substrates (330) may be placed vertically relative to the ground such that fluid is configured to flow downward through spaces between the substrates.

[0138] In some embodiments, the fluid diffuser may be configured to regulate the fluid flow rate at one or more substrates of the device. For example, the fluid diffuser may be configured to receive a fluid and output a substantially uniform and laminar flow to each substrate of a plurality of substrates. FIG. 4A is a perspective view of a fluid diffuser (400) having a first portion (420) and a second portion (430). The fluid diffuser comprises a plurality of openings (410). The fluid diffuser (400) has a boundary (440) of the first portion (420) and the second portion (430). The first portion (420) and the second portion (430) may be curved relative to one another at the boundary (440). In some of these embodiments, the boundary (440) may include one or more of the plurality of openings (412). The boundary (440) may be linear or non-linear.

[0139] The first portion (420) is angled relative to the second portion (430) at an angle between about 90 degrees and about 170 degrees, including all values ​​and subranges therebetween. For example, Figure 4D shows a side view of the fluid diffuser (400) in which the first portion (420) is angled at about 150 degrees relative to the second portion (430). A larger angle between the first portion (420) and the second portion (430) allows for a lower height of the fluid diffuser (400), which in turn reduces the volume of the empty space between the inlet and the plurality of substrates.

[0140] The first portion (420) and the second portion (430) can be angled at up to about 80 degrees relative to the plurality of substrates. In some embodiments, the plurality of openings are at least about 700 μm apart. 2 For example, each opening has a diameter of about 30 μm to allow the growth medium to pass through the fluid diffuser (400).

[0141] FIG. 4B is a plan view of the fluid diffuser (400). In some embodiments, the plurality of openings are substantially equally spaced apart. In some embodiments, each opening (410, 412) of the plurality of openings has a diameter of at least about 30 μm. For example, the diameter of the openings (410, 412) can range from about 30 μm to about 10 cm, and from about 0.5 cm to about 3 cm, including all values ​​and subranges therebetween. FIG. 4C is another perspective view of the fluid diffuser (400). As shown in FIG. 4D, the first portion (420) and the second portion (430) are substantially flat with minimal thickness. The configuration of the fluid diffuser (400) can be configured to output a substantially uniform laminar flow to a plurality of substrates while reducing fouling due to material build-up and clogging, and facilitating sterilization. The shape of the openings (410, 412) is circular in FIGS. 4A-4D, but may be any shape.

[0142] The amount of meat product prepared by the apparatus described herein corresponds to the number and surface area of ​​the substrates of the apparatus. For example, multiple parallel substrates can be configured to produce commercial-scale quantities of edible meat products. Figures 5A and 5B are perspective and side views, respectively, of multiple substrates (500). In some embodiments, the apparatus for preparing meat products includes one or more substrates (500) configured to culture one or more cells on its surface to form a meat product. In some embodiments, the one or more substrates (500) are removable from the apparatus so that they can be washed or conserved and reused. As shown in Figure 5A, the multiple substrates (500) are planar and substantially parallel to each other. In some embodiments, the apparatus has a single substrate or multiple substantially parallel substrates (e.g., parallel plate configuration). The parallel plate configuration can increase the surface area of ​​the attached tissue sheet. In some embodiments, the parallel plates are aligned with the direction of fluid flow. For example, the plates and corresponding fluid channels are positioned perpendicular to the ground.

[0143] In some embodiments, the device may have multiple fluid channels (510). For example, the multiple substrates (500) may define multiple fluid channels (510). Each fluid channel (510) may be associated with at least one of the multiple substrates (500) in that the fluid channel (510) includes a space between adjacent substrates (500). If the multiple substrates (500) are planar, the fluid channel (510) may be planar. As shown in FIG. 5B, the fluid may be configured to flow unidirectionally through the multiple fluid channels (510). For example, the multiple planar substrates may be separated by a predetermined gap (e.g., a vertical gap if the substrates are vertical to the ground) through which the fluid may be perfused so that meat products may grow in the multiple substrates. In some embodiments, the multiple fluid channels are substantially parallel to one another. In some embodiments, each fluid channel may have a width between about 0.3 mm and about 5.0 cm, including all values ​​and subranges therebetween.

[0144] The plurality of substrates (500) may allow for high density growth of meat products. For example, the substrates (500) may be configured to grow meat products on both sides of the substrate. In some embodiments, each substrate (500) has dimensions having a width between about 10 cm and about 400 cm, and a length between about 10 cm and about 200 cm. In some embodiments, the plurality of substrates (500) may include a number of substrates up to about 10,000,000, including all values ​​and subranges therebetween.

[0145] In some embodiments where the substrate has a spiral (e.g., coil) shape, the substrate has a thickness between about 0.1 mm and about 10 mm, a length between about 10 cm and about 300 cm, a width between about 10 cm and about 200 mm, and a spacing between adjacent spirals of about 1 mm and about 10 mm, including all values ​​and subranges therebetween. For example, a spiral substrate may have a thickness of about 2 mm, a width of about 128 mm, and a spacing between adjacent spirals of about 3 mm. In some embodiments, the spacing between adjacent (e.g., adjacent) substrates may be between about 0.3 mm and about 5.0 cm, including all values ​​and subranges therebetween. In some embodiments, each substrate may have a length of about 430 cm, including all values ​​and subranges therebetween. 2 From about 100,000,000 cm 2 The area is between .

[0146] 9A and 9B are cross-sectional side views of fluid flow vector diagrams (900, 960) through an apparatus for preparing meat products as described herein. FIG. 9A shows the fluid flow rate (900) of a cross-sectional image of an apparatus (910) having a housing (920), a fluid diffuser (930), a plurality of substrates (940), and a collector (950). Fluid enters the apparatus (910) from the top of the housing (920) in a relatively high velocity, non-uniform, non-laminar flow. The fluid diffuser (930) receives the fluid and diffuses (e.g., regulates, modulates) the flow such that the plurality of substrates (940) receive a substantially uniform laminar fluid flow, thereby enabling generally uniform cell growth in each of the plurality of substrates. FIG. 9B is a detailed cross-sectional image of the fluid flow rate (960) of the plurality of substrates, showing the substantially uniform laminar fluid flow through the substrates. The fluid flow is generally regular and symmetrically distributed throughout the plurality of substrates (940). The fluid passes through fluid channels between adjacent substrates to the collector (950) and an outlet of the housing (920). The collector (950) can be configured to maintain a flow rate sufficient to reduce back pressure within the device (910). For example, the collector (950) can be configured to maintain a flow rate sufficient to prevent fluid from backflowing onto multiple substrates.

[0147] 6A-6C are exemplary diagrams of an exemplary embodiment of a collector (600) including one or more openings (610) and a connector (620) configured to detachably engage the collector (600) to an apparatus. In some embodiments, the collector (600) may be configured to receive fluid and meat products (e.g., one or more cell sheets) from one or more substrates. Fluid passes through the collector (600) and exits the apparatus through an outlet while the meat products are held in the collector (600) until harvested. For example, the collector (600) may be configured to receive meat products grown on a substrate after the meat products are separated from the substrate using a fluid flow. In some embodiments, the meat products may fall into the collector (600) by gravity. For example, the collector (600) may be placed below one or more substrates such that the collector (600) captures and holds the falling meat products.

[0148] In some embodiments, the collector (600) may be disposed within a cavity of the device. Additionally or alternatively, the collector may be bonded to the exterior of the device. In some embodiments, the collector (600) may be at least about 700 μm 2 For example, each opening may include a diameter between about 30 μm and about 2.5 cm, including all values ​​and subranges therebetween. In some embodiments, the collector (600) may be a separate component from the housing or may be integrated with the housing. In some embodiments where the collector (600) is external to the housing, the collector may have a diameter of at least about 20 μm. 2 The aperture may have one or more openings having an area of

[0149] 7A, 7B, 7C, and 7D are exemplary perspective views of an assembly process of an apparatus (700) for preparing meat products. In some embodiments, a plurality of substrates (730) coupled to a holder (750) may be inserted into the housing (710). The axis of rotation (712) of the housing (710) is shown in FIGS. 7A and 7B. As described in more detail herein, the housing (710) may be rotated about the axis of rotation (712) to assist in seeding the plurality of substrates (730). FIG. 7B shows the plurality of substrates (730) placed within a cavity of the housing (710). FIG. 7C shows a fluid diffuser (720) inserted into the cavity (710) above the plurality of substrates (730). The housing (710) may further include a lid (not shown) configured to seal an open end of the housing (710) to promote a sterile environment within the apparatus (700). Once the device (700) is assembled, a plurality of cells are provided on a plurality of substrates (730).

[0150] In some variations, a rotor (e.g., a rotation mechanism) coupled to the housing may be configured to rotate the device to aid in cell seeding. Figures 8A, 8B, and 8C are exemplary side views of an exemplary embodiment of an apparatus (850) or a rotating process (800) for preparing meat products. The apparatus may rotate about a rotation axis (810) having a rotor (not shown). The substrate (810) may be disposed vertically within the housing (850) along the length of the housing (850). As the apparatus (850) may rotate, the substrate (810) within the housing (850) may rotate about the rotation axis (810). As shown in Figures 8A and 8C, the rotation of the apparatus (850) about the rotation axis (810) allows either side of the substrate (820) to be held upward, thereby allowing cell sedimentation by gravity. The device (850) can be rotated upright as shown in FIG. 8B to position the substrate (820) perpendicular to the ground and allow fluid to flow from the top to the bottom of the device (850).

[0151] 10A-12D show a second exemplary embodiment of an apparatus for preparing a meat product. FIG. 10A is a perspective view of an embodiment of an apparatus (1000) for preparing a meat product, without displaying the housing (for clarity). FIG. 10B is a front view of the apparatus (1000), and FIG. 10C is a side view of the internal components of the apparatus (1000). The apparatus (1000) shown in FIG. 10A has a fluid diffuser (1010), a plurality of substrates (1020), and a holder (1030). In some embodiments, the holder (1030) can be configured to detachably engage (e.g., hold) one or more of the fluid diffusers (1010) and the plurality of substrates (1020). In some embodiments, the holder (1030) can be integrated into an inner wall of the housing of the apparatus. The fluid diffuser (1010) may be disposed on a proximal side (e.g., on an upper surface) of the plurality of substrates (1020), and a collector (not shown) may be disposed on a distal side (e.g., on a lower surface) of the plurality of substrates (1020). Any of the fluid diffuser (1010), substrates (1020), and holder (1030) may be removable from the housing or fixed to the housing. In some embodiments, the fluid (e.g., growth medium) may be configured to flow from the fluid diffuser (1010) in a direction (e.g., from proximal to distal, top to bottom) to the substrates (1020).

[0152] The fluid diffuser (1010) may be configured to generally distribute the received fluid flow to each of the substrates (1020). The fluid diffuser (1010) may be coupled to a proximal end of the substrates (1020). In some embodiments, the fluid diffuser (1010) may be slidably and releasably engaged to the holder (1030) without tools or fasteners, which facilitates maintenance and sterilization of the device. The collector (not shown) may be configured to receive the meat product and fluid from each of the substrates (1020). In some embodiments, the collector may be coupled to a distal end of the substrates (1020). Additionally or alternatively, the collector may be located outside the device (1000). For example, the collector may receive the output of the housing.

[0153] In some embodiments, the plurality of substrates (1010) are configured to releasably engage with the holder (1030). For example, the plurality of substrates (1020) may be slidably and releasably engaged with the holder (1030) without tools or fasteners, which allows for easy maintenance and sterilization of the plurality of substrates (1020). In some embodiments, the plurality of substrates (1020) may be disposed in a substantially vertical orientation relative to the ground such that fluid may be configured to flow downward through spaces between the substrates. In some embodiments, the plurality of substrates (1020) may be disposed in a substantially horizontal orientation relative to the ground during one or more steps of the method of preparing a meat product. For example, the meat product may be grown on the plurality of substrates (1020) in a substantially horizontal orientation for one or more days.

[0154] 11A is a perspective view of a fluid diffuser (1100) including a first portion (1120), a second portion (1122), a third portion (1124), and a fourth portion (1126). The fluid diffuser (1100) may comprise a plurality of openings (1110). In some embodiments, the plurality of openings is at least about 700 μm 2 For example, each opening may have a diameter of about 30 μm to allow the growth medium to pass through the fluid diffuser (1100). The fluid diffuser (1100) may include a boundary (1130) between adjacent portions where the portions are curved relative to one another. In some of these embodiments, the boundary (1130) may include one or more of the plurality of openings (1132). The boundary (1130) may be linear or non-linear. In some embodiments, the fluid diffuser (1100) may further include a connector (1140).

[0155] FIG. 11B is a side view of the fluid diffuser (1100). The opposing portions can be angled relative to one another between about 90 degrees and about 170 degrees, including all values ​​and subranges therebetween. For example, the first portion (1120) can be angled at about 150 degrees relative to the third portion (1124), and the second portion (1122) can be angled relative to the fourth portion (1126). A larger angle between the portions can allow the height of the fluid diffuser (1100) to be lower, thereby reducing the volume of the empty space between the inlet and the substrates. Each portion of the fluid diffuser (1100) can be angled at up to about 80 degrees relative to the substrates.

[0156] FIG. 11C is a plan view of the fluid diffuser (1100). In some embodiments, the plurality of openings are substantially equally spaced apart. In some embodiments, each opening (1110, 1132) of the plurality of openings has a diameter of at least about 30 μm. For example, the diameter of the openings (1110, 1132) can be between about 30 μm and about 10 cm, between about 0.5 cm and about 3 cm, including all values ​​and subranges therebetween. The configuration of the fluid diffuser (1100) can output a substantially uniform laminar flow to a plurality of substrates while reducing fouling due to material build-up and clogging, and facilitating sterilization. The shape of the openings (1110, 1132) is shown as a circle in FIGS. 11A-11C, but may be any shape.

[0157] FIG. 12A is a perspective view of the substrates (1200). FIG. 12B is a side view of the substrates (1200). The substrates (1200) may collectively form a generally cylindrical shape that fits within a corresponding generally cylindrical housing (not shown). In some embodiments, each substrate has substantially the same thickness and height as shown in FIG. 12C, while having different widths as shown in FIGS. 12A and 12B. In some embodiments, the fluid channels (1210) include spaces between adjacent substrates (1200). For example, the fluid channels (1210) may have a width between about 0.3 mm and about 5.0 cm, including all values ​​and subranges therebetween. The fluid channels (1210) are planar and coincide with the plane of the substrates (1200).

[0158] In some embodiments, one or more substrates (1200) are removable from the device so that they can be cleaned, maintained, and reused. As shown in FIG. 12C, the substrates (1200) are planar and substantially parallel to one another. In some embodiments, the device includes a single substrate or multiple substantially parallel substrates (e.g., parallel plate configuration). In some embodiments, the parallel plates are aligned with the direction of fluid flow. For example, the plates and corresponding fluid channels are positioned perpendicular to the ground. The multiple planar substrates can be separated by a predetermined gap (e.g., a vertical gap if the substrates are perpendicular to the ground) through which fluid can be perfused so that meat products can grow in the multiple substrates. In some embodiments, the multiple fluid channels are substantially parallel to one another. In some embodiments, each fluid channel can have a width between about 0.3 mm and about 5.0 cm.

[0159] The plurality of substrates (1200) may allow for high density growth of meat products. For example, the substrates (1200) are configured to grow meat products on both sides of the substrate. In some embodiments, each substrate (1200) has dimensions of between about 10 cm and about 400 cm in width, between about 10 cm and about 200 cm in length, including all values ​​and subranges therebetween. In some embodiments, the plurality of substrates (1200) may include up to about 1000 substrates, including all values ​​and subranges therebetween. In some embodiments, the spacing between adjacent (e.g., adjacent) substrates may be between about 0.3 mm and about 5.0 cm, including all values ​​and subranges therebetween. In some embodiments, each substrate may be up to about 140,000 cm in width, including all values ​​and subranges therebetween. 2 In some embodiments, each substrate has an area of ​​up to about 4,000 cm 2 In some embodiments, each substrate has an area of ​​about 430 cm 2 From about 8,000cm 2 The area is between .

[0160] 13A-17C show a third exemplary embodiment of an apparatus for preparing a meat product. FIG. 13A is a cross-sectional side view of an apparatus (1300) for preparing a meat product. The apparatus (1300) may include a housing (1310) including an inlet (1312), an outlet (1314), and further defining a cavity (1316). Within the cavity (1316) may be located a fluid diffuser (1320), at least one substrate (1330), and a holder (1350). In some embodiments, the holder (1350) may be configured to releasably engage (e.g., hold) the one or more fluid diffusers (1320) and the at least one substrate (1330). The holder (1350) may be coupled to an inner wall of the housing (1310). In some embodiments, the holder (1350) may be integral to an inner wall of the housing (1310). The fluid diffuser (1320) may be disposed between the inlet (1312) and at least one substrate (1330). For example, the fluid diffuser (1320) may be disposed within a headspace (e.g., an empty space above a substrate) of the housing (1310). Either the fluid diffuser (1320) or the substrate (1330) may be removable from the housing (1310) or may be fixed to the housing (1310). In some embodiments, the fluid (e.g., liquid, growth medium, gas) may be configured to flow in a direction from the inlet (1312) to the outlet (1314). For example, the fluid may flow from a higher position (e.g., the inlet (1312)) to a lower position (e.g., the outlet (1314)). In some embodiments, the fluid (e.g., the growth medium) may be configured to flow from the outlet (1314) to the inlet (1312). In some embodiments, fluid flow may be configured to change direction during use through inlet (1312) and outlet (1314). In some embodiments, housing (1310) may include multiple inlets and outlets (e.g., growth medium inlet, gas inlet, gas outlet).

[0161] FIG. 13A is a cross-sectional side view of an apparatus (1300) including a rolled (e.g., spiral-shaped) substrate (1330). FIG. 13B is another cross-sectional side view of the apparatus (1300) shown in FIG. 13A, where the substrate (1330) can be rolled to have multiple turns, as described in more detail with reference to FIGS. 16A-16F. FIG. 13C is a perspective view of the apparatus (1300). In some embodiments, the apparatus can include multiple fluid diffusers (1320) disposed in a head space of the housing (1310). For example, the fluid diffusers (1310) can be positioned at a predetermined distance away from the substrate (1330). In some embodiments, the fluid diffusers (1320) can intersect with a central longitudinal axis of the housing (1310). The fluid diffusers (1320) can be configured to generally distribute a received fluid flow to one or more substrates (1330).

[0162] In some embodiments, the one or more substrates (1330) may be configured to fill a predetermined volume of the housing (1310). For example, the one or more substrates (1330) may extend radially from the central longitudinal axis of the housing (1310) to the holder (1350) (e.g., the inner periphery) to increase the volume of tissue grown within the housing (1310). In some embodiments, the substrate (1330) may include one or more spacing features (e.g., spacers, protrusions) configured to maintain a predetermined distance between substrate surfaces (e.g., different peripheries of the substrate, different parallel plates). For example, as shown in more detail with reference to Figures 16F and 16G, the substrate may include spaced protrusions configured to contact another portion of the substrate or maintain a predetermined distance between substrate surfaces.

[0163] 13D and 13E are cross-sectional perspective views of the device (1300). In some embodiments, the fluid may be configured to flow downward from the fluid diffuser through a space between the circumference of the substrate (1330). For example, the scroll-like substrate (1330) may define a plurality of parallel fluid channels. In some embodiments, the fluid diffuser (1320) may be coupled to a proximal end of one or more substrates (1330). In some embodiments, the fluid diffuser (1320) may be configured to detachably engage a spacer (1354) of the holder (1350) such that at least a portion of the fluid diffuser (1320) is disposed within the cavity (1316). For example, the fluid diffuser (1320) may be slidably and detachably engaged with the holder (1350) to facilitate maintenance and sterilization of the device (1300). In some embodiments, a collector (not shown) may be coupled to a distal end of the plurality of substrates (1330). In some embodiments, one or more substrates (1330) can be configured to releasably engage the housing (1330). For example, one or more substrates (1330) can be slidably and releasably engaged with the holder (1350) to facilitate maintenance and sterilization of the one or more substrates (1330). In some embodiments, the longitudinal axis of the substrate (1330) can be parallel to the longitudinal axis of the housing (1310).

[0164] 13F is a top view of the apparatus (1300). In some embodiments, the fluid diffuser (1320) can have a diameter smaller than the diameter of the one or more substrates (1330) and the holder (1350). In some embodiments, the fluid diffuser (1320) can be configured to regulate a fluid flow rate over the one or more substrates (1330) of the apparatus (1310). For example, the fluid diffuser (1320) can be configured to receive a fluid and output a substantially uniform laminar flow of fluid to each of the multiple substrates.

[0165] In some embodiments, the fluid diffuser (1320) may be coupled to the housing and / or one or more substrates. The fluid diffuser (1320) may be configured to distribute fluid to the substrate in a predetermined flow pattern. For example, the fluid diffuser may be configured to allow a substantially uniform or laminar flow across a diameter of the substrate. As described in more detail herein, the fluid diffuser may have a tapered surface. In some embodiments, the ratio of the dimensions (e.g., diameter) of the fluid diffuser to the dimensions of the substrate is between about 3:4 and about 1:1, between about 1,000:1 and about 1:1, including all subranges therebetween. Additionally or alternatively, the fluid diffuser may comprise one or more openings. For example, the multiple openings may be substantially equally spaced apart.

[0166] In some embodiments, the fluid diffuser can be radially symmetric. In some embodiments, the cavity includes a head space, and the fluid diffuser is disposed within the head space of the housing. In some embodiments, the longitudinal axis of the fluid diffuser can be substantially perpendicular to the substrate. In some embodiments, the fluid diffuser and the substrate have a predetermined distance. In some embodiments, the predetermined distance is up to about 20 cm.

[0167] In some embodiments, the fluid diffuser may be radially symmetric. In some embodiments, the cavity includes a headspace, and the fluid diffuser is disposed within the headspace of the housing. In some embodiments, the longitudinal axis of the fluid diffuser may be substantially perpendicular to the substrate. In some embodiments, the fluid diffuser and the substrate have a predetermined spacing. In some embodiments, the predetermined spacing may be up to about 20 cm. In some embodiments, the fluid diffuser may include polytetrafluoroethylene (PTFE).

[0168] FIG. 14A is a plan view of a flat plate fluid diffuser (1400) having a flat disk shape. The fluid diffuser (1400) may have a diameter smaller than the circumference of the inner diameter of the housing. FIG. 14B is a perspective view of the fluid diffuser (1400) shown in FIG. 14A. In some embodiments, the fluid diffuser may have one or more openings (see FIG. 15D and FIG. 18B) configured to allow liquid (e.g., growth medium) to pass through the fluid diffuser (1400). That is, the openings may have a length through the thickness of the fluid diffuser. The openings may be configured to distribute the fluid flow more evenly to one or more substrates placed under the fluid diffuser. For example, each opening may have a diameter of about 30 μm, thereby allowing the growth medium to pass through the fluid diffuser. In some embodiments, the fluid diffuser (1400) may be perpendicular to one or more substrates.

[0169] 14C and 14D are side and perspective views, respectively, of the fluid diffuser (1400) and holder (1410). In some embodiments, the holder (1410) may be located along the central longitudinal axis of the housing (not shown for clarity). The configuration of the fluid diffuser (1400) may output a substantially uniform laminar flow to multiple substrates while reducing fouling due to material build-up and clogging, and facilitating sterilization. For example, fluid may be configured to flow over the top and bottom surfaces of the fluid diffuser (1400). The shape of the fluid diffuser (1400) is circular in FIGS. 14A-14D, but may be any shape. In some variations, the fluid diffuser and substrate may be separated by up to about 20 cm.

[0170] 15-15C are perspective views of variations of angled fluid diffusers (1500, 1510, 1520) having a bicone shape. A bicone (e.g., a bicone) refers to a three-dimensional surface obtained by rotating a rhombus around one of its axes of symmetry. That is, a bicone may define a structure generated by joining two congruent right circular cones at their bases. In some embodiments, a bicone may have circular and orthogonal symmetry. The fluid diffusers (1500, 1510, 1520) shown in FIGS. 15A-15C show different heights and diameters. The fluid diffusers (1500, 1510, 1520) may have a diameter smaller than the circumference of the inner diameter of the housing. In some variations, the bicone may include a single part or multiple parts.

[0171] In some embodiments, the fluid diffuser (1500, 1510, 1520) can have a cone angle between about 90 degrees and about 170 degrees. Each cone of the bi-cone can have the same or different cone angle. That is, each cone of the bi-cone can have the same or different height.

[0172] The larger the cone, the lower the height of the fluid diffuser (400) can be, thereby reducing the volume of the empty space between the inlet and the substrates.

[0173] In some embodiments, the configuration of the openings in the fluid diffuser may have a symmetrical pattern. For example, each opening may have substantially the same diameter and the spacing between adjacent openings may be substantially the same. In some embodiments, the openings may extend across substantially the entire side of the fluid diffuser.

[0174] FIG. 15B shows an example of a fluid flow (1550) through a fluid diffuser (1510). The fluid can be configured to flow over substantially the entire surface area of ​​the fluid diffuser (1510). That is, the fluid can be configured to flow over the top and bottom surfaces of a biconical fluid diffuser. When the fluid flows over the peripheral (e.g., circumferential) edge of the fluid diffuser, the fluid flows along the underside of the fluid diffuser due to adhesion forces. Adhesion forces are intermolecular attractive forces between different types or states of molecules. An example of adhesion forces is the phenomenon of water wetting a solid surface. Wetting occurs due to intermolecular forces between water and a solid surface. However, as the fluid flows over the underside, the fluid falls off the fluid diffuser due to gravity and may fall radially larger than the diameter of the substrate. For example, the fluid may flow over the substrate in a radially symmetric manner. A radially symmetric fluid diffuser allows the fluid to flow in substantially equal paths, which can provide a substantially uniform distribution of the fluid to one or more substrates.

[0175] In some embodiments, an apparatus for preparing a meat product may include a housing defining a cavity, a substrate disposed within the cavity and configured to support growth of the meat product, and a fluid diffuser having a tapered surface and configured to distribute a fluid to the substrate in a predetermined flow pattern.

[0176] In some embodiments, the tapered surface may include one or more of a concave, conical, frustum, step, and flare shape. In some embodiments, the tapered surface may be angled up to about 80 degrees relative to the substrate. In some embodiments, the fluid diffuser may have a bi-cone.

[0177] In some embodiments, the bicone may include a first surface and a second surface opposite the first surface. The first surface may be configured to receive a fluid, and the second surface may be configured to receive the fluid from the first surface and distribute the fluid to the substrate. In some embodiments, the fluid diffuser may be configured to allow a substantially uniform or laminar flow across a diameter of the substrate. In some embodiments, the fluid diffuser may include one or more openings. In some embodiments, the one or more openings may include a plurality of openings that are substantially equally spaced apart. In some embodiments, the fluid diffuser is radially symmetric. In some embodiments, the cavity may include a headspace, and the fluid diffuser may be located within the headspace of the housing. In some embodiments, the longitudinal axis of the fluid diffuser may be substantially perpendicular to the substrate. In some embodiments, the fluid diffuser and the substrate may have a predetermined spacing. In some embodiments, the predetermined spacing may be up to about 20 cm.

[0178] FIG. 15D is a plan view of a fluid diffuser (1530) having a conical shape (e.g., a single cone). The fluid diffuser (1530) may include one or more openings (1532) configured to allow a liquid (e.g., growth medium) to pass through the fluid diffuser (1530). For example, the openings may be substantially equally spaced. In some embodiments, each opening (1532) may have a predetermined diameter. For example, the diameters of the openings (1532) may be the same or may vary as a function of distance from the center of the fluid diffuser (1530). In some embodiments, the fluid diffuser (1500) may be perpendicular to one or more substrates. For example, the fluid may be configured to flow over the top surface and through the openings (1532) of the fluid diffuser (1500).

[0179] The amount of meat product prepared by the apparatus described herein corresponds to the number and surface area of ​​the substrates in the apparatus. For example, one or more substrates can be configured to produce commercial-scale quantities of edible meat product.

[0180] FIG. 16A is a perspective view of the substrate (1600). FIG. 16B is a side view of the substrate (1600). FIG. 16C is a plan view of the substrate (1600), and FIG. 16D is a detailed plan view. FIG. 16E is a plan view of the substrate installed in a housing. FIG. 16F is a schematic diagram of a spacer of the substrate. In some embodiments, the apparatus for preparing a meat product may include one or more substrates (1600) configured to promote the cultivation of one or more cells on its surface to form a meat product. In some embodiments, the one or more substrates (1600) may be removable from the corresponding apparatus so that they may be cleaned or conserved and reused. Additionally or alternatively, the one or more substrates may be sterilized within the apparatus itself. In some embodiments, the one or more substrates (1600) may be planar and substantially parallel to one another. In some embodiments, the apparatus may include a single substrate or multiple substrates. In some embodiments, the substrates may be configured to releasably engage the housing. For example, multiple substrates may be joined (e.g., attached) to one another in one or more configurations (e.g., end-to-end, spaced apart, overlapping). One or more fasteners may be used to join adjacent substrates to one another.

[0181] In some embodiments, the substrate (1600) may have a scroll-like configuration having multiple (e.g., 2, 3, 4, 5, or more) layers (e.g., revolutions) wound (e.g., wrapped around) a longitudinal axis (e.g., a central axis). In the case of a scroll-like configuration, the substrate (1600) may have a generally cylindrical shape with a circular or elliptical cross-sectional shape. For example, the scroll-like substrate (1600) may comprise an elliptical cylinder (e.g., a cylinder having an elliptical cross-section).

[0182] The substrate (1600) may have a configuration that is wound around a given axis two or more times. That is, the substrate (1600) may be bent more than 360 degrees around the given axis. For example, the substrate may have multiple turns around the given axis. In some embodiments, the substrate (1600) may be bent around a longitudinal axis of the housing. For example, the substrate (1600) may be disposed (e.g., installed) within a cavity of the housing and aligned along a central longitudinal axis of the housing.

[0183] In some embodiments, the substrate (1600) has an inner end (1610) (e.g., the innermost portion of the substrate in a roll, the proximal end) and an outer end (1620) (e.g., the outermost portion of the substrate in a roll, the distal end). For example, FIG. 16D shows the inner end (1610) of the substrate (1600), which may define the beginning or center of the substrate (1600), and FIG. 16A shows the outer end (1620) of the substrate (1600), which may define the end or last layer of the substrate (1600). The substrate (1600) in a roll can have one or more revolutions or turns such that the substrates overlap each other. The substrates (1600) in a roll described herein can provide increased surface area for the support and growth of the adherent tissue sheet. Additionally, the substrate (1600) in a roll can have a shape that is more rigid than a flat substrate (e.g., a plate). In some embodiments, the substrate (1600) can have an outer diameter of at least about 5 cm. In some embodiments, the substrate (1600) can have a height of up to about 20 cm, 50 cm, 80 cm, about 100 cm, including all values ​​and subranges therebetween.

[0184] In some embodiments, the substrate can be manufactured by stamping one or more protrusions onto a sheet of thin metal having spring elasticity. The substrate can be configured to be rolled up and placed within the housing, where it naturally expands within the housing, moving away from one another and filling the cavity of the housing. In some embodiments, the substrate can be removably coupled to the housing, i.e., the substrate does not have to be fixed or attached to the housing.

[0185] In some embodiments, the scrolled substrate may be aligned along the fluid flow. For example, the substrate and corresponding fluid channels may be configured to be at an angle (e.g., vertical, off-axis) with respect to the ground. In some embodiments, the device may include multiple fluid channels (1660). For example, the scrolled substrate (16000) may define multiple fluid channels (1660) between adjacent peripheries (e.g., in the space between adjacent layers of the substrate (1600)). Each fluid channel (1660) may be associated with at least one turn of the substrate (16000) in that the fluid channels (1660) include a space between successive peripheries. For example, adjacent peripheries of the substrate (1600) may have a predetermined spacing. The fluid may be configured to flow in one direction (e.g., from a higher position to a lower position) through the multiple fluid channels (1660). For example, each layer of substrate (1600) may be separated by a predetermined distance (e.g., a gap) through which a fluid may be perfused so that meat products may grow on the substrate (1600). In some embodiments, the plurality of fluid channels (1660) are substantially curved (e.g., concentric) and parallel to one another. In some embodiments, each fluid channel (1660) may have a width between about 0.3 mm and about 5.0 cm, including all values ​​and subranges therebetween.

[0186] In some embodiments, the apparatus for preparing a meat product may include a housing as described herein and a substrate (1600) disposed within a cavity of the housing, the substrate (1600) being spiral-shaped and having a surface configured to encourage growth of meat products. For example, the substrate (1600) may allow for high density growth of meat products. In some embodiments, the substrate (1600) may be configured to grow meat products on one or both sides of the substrate. In some embodiments, the substrate (1600) may include dimensions of between about 10 cm and about 300 cm in width, between about 10 cm and about 200 m in length, including all values ​​and subranges therebetween. In some embodiments, the plurality of substrates (1600) may include up to about 10,000,000 substrates, including all values ​​and subranges therebetween. In some embodiments, the substrate may include one or more of stainless steel, ceramic, and polymer.

[0187] In some embodiments, the housing may include a plurality of scroll-like (e.g., spiral-like) substrates (1600), each wound about its own axis. For example, a scroll-like main substrate having a first diameter may be located in the center of the housing, and scroll-like minor substrates having a second diameter smaller than the first diameter may be located around the circumference of the scroll-like main substrate. As another example, the scroll-like substrates may be arranged in parallel lines within the cavity of the housing. That is, the axes of each scroll-like substrate may be non-intersecting and parallel.

[0188] In some embodiments, the substrate (1600) may have a thickness of at least about 1 μm. The stiffness of the scrolled substrate (1600) due to its spiral shape may allow for a corresponding reduction in thickness of the substrate (1600), thereby allowing for additional circumferences of the substrate within a given volume. In some embodiments, each circumference of the substrate may be separated by a predetermined distance configured to promote growth of the edible meat product and allow fluid to flow through the substrate (1600). For example, the predetermined distance between adjacent circumferences of the substrate may be at least about 0.8 mm.

[0189] In some embodiments, a spiral may include multiple substrates. For example, the substrate may include a first substrate portion and a second substrate portion formed separately from the first substrate portion. An end of the first substrate portion may be proximate to an end of the second substrate portion. In some embodiments, an end of the first substrate portion may be joined to an end of the second substrate portion. For example, an end of the first substrate portion overlaps an end of the second substrate portion. Alternatively, the stop may be configured such that the ends of each substrate portion are joined together without the portions contacting each other.

[0190] In some embodiments, the substrate may have one or more features (e.g., spacers, protrusions) configured to maintain a predetermined spacing between adjacent layers of the substrate. In some embodiments, the substrate may include a plurality of protrusions configured to space the substrate in a roll disposed within the housing. In some embodiments, the one or more protrusions may have an approximately linear shape. The one or more protrusions may be imprinted on the surface of the substrate. The protrusions may be configured to allow the tissue sheet to grow between the spaced apart protrusions. For example, protrusions having a linear shape may promote the growth of a tissue sheet with a corresponding linear edge. In some embodiments, linear protrusions may reduce tears and / or damage to the tissue and aid in the separation of the tissue sheet from the substrate. In some embodiments, the one or more protrusions may have a height of at least about 0.8 mm. Adjacent protrusions may be spaced at a predetermined spacing along the surface of the substrate, including between about 2 cm and about 8 cm, and all values ​​and subranges therebetween.

[0191] 16F and 16G are plan views of the surfaces of substrates (1600, 1602) having spacers (1630, 1632). In some embodiments, the substrates (1600, 1602) can have an inner end (1610) and an outer end (1620). The spacers (1630) can have one or more elongated, rounded shapes. Additionally or alternatively, the spacers (1632) can be one or more of a generally circular, spherical, cylindrical, hemispherical, ovoid, elliptical, combinations thereof, and the like.

[0192] In some embodiments, at least one of the substrate and the housing may be angled with respect to the ground to promote cell attachment and tissue growth on the substrate. For example, the housing may be configured to be placed on the ground and the substrate of the housing may define a longitudinal axis that is at an acute angle with the ground. FIG. 2E shows a housing (260) angled with respect to the longitudinal axis (290). Similarly, the substrate (not shown) within the housing (260) may be angled with respect to the longitudinal axis (290). In some embodiments, the acute angle may be at least about 85 degrees. Thus, the surface of the substrate facing away from the ground may be configured to promote the growth of meat products. In some embodiments, a slightly angled substrate may promote laminar flow while promoting cell attachment and growth on the substrate.

[0193] 18A-18E are vector diagrams (1800, 1802, 1804, 1806, 1808) of fluid flow through an apparatus for preparing a meat product as described herein. FIG. 18A shows fluid flow rates (1800) in a cross-sectional image of an apparatus including a fluid diffuser (1810) and a scrolled substrate (1820). A non-uniform, non-laminar flow (1850) of fluid enters the apparatus through an inlet (1802) at a relatively high velocity. The fluid diffuser (1810) receives the fluid and diffuses (e.g., regulates, modulates) the fluid flow (1852) such that the scrolled substrate (1820) receives a substantially uniform, laminar flow of fluid (1854), thereby enabling consistent cell growth throughout each layer (e.g., circumference) of the substrate (1820). The fluid flow may be generally regular and symmetrically distributed throughout the layers of the substrate (1820). For example, Figure 18C shows a perspective view of a fluid flow (1804) over a cone-shaped fluid diffuser (1810) and through a scroll-like substrate (1820). Returning to Figure 18A, the fluid (1856) passes through fluid channels between adjacent layers of the substrate (1820) and then through an outlet (1856) of the housing (1820). In some embodiments, the system can be configured to maintain a sufficient flow rate to reduce backpressure within the device.

[0194] FIG. 18B is a detailed cross-sectional view of the fluid flow rate (1802) of a fluid diffuser (1810) having a scroll-like substrate and multiple openings (1812). FIG. 18D is another cross-sectional image of the fluid flow rate (1806) of a biconical fluid diffuser (1811). The fluid diffuser (1811) is configured to allow fluid flow (1850) over a first tapered portion (e.g., upper cone) (1813) and fluid flow (1852) over a second tapered portion (e.g., lower cone) (1815). For example, the fluid (1852) can flow from the first tapered portion (1813) to the second tapered portion (1815) due to viscosity. As the fluid flows under the biconical diffuser (1811) (e.g., second tapered portion (1815)), the fluid can fall to the substrate (1820) due to gravity. In some embodiments, the fluid may flow through the substrate (1820) with radial symmetry. Figure 18E is a detailed cross-sectional image of the fluid flow velocity (1808) through the substrate (1820).

[0195] III. method Also described herein are methods for preparing the meat products described herein. The methods described herein allow for the production and separation of meat products within the apparatus. In some embodiments, a quantity of cells are provided to a substrate, which allows the cells to grow into a meat product (e.g., a cell sheet). Once the cells meet predetermined growth requirements, such as size and maturity, the meat product may be separated from the substrate. For example, the meat product may be separated from the substrate in a substantially intact and / or simultaneous manner using a fluid flow. The apparatus described herein for producing meat products may be further sterilized for reuse.

[0196] Generally, the methods described herein include contacting a substrate with a quantity of cells for a predetermined period of time. Growth medium may be delivered to the cells to promote cell growth, cell differentiation, and / or maintenance of an undifferentiated cell state. For example, a quantity of cells may be grown on the surface of any of the substrates described herein. The cells mechanically interact and grow on the surface of the substrate to form a continuous cell sheet. Multiple substrates may allow for the production and collection of multiple cell sheets. The cells grown on the substrate may be separated as an unbroken (e.g., continuous) meat product (e.g., cell sheet) and / or in a predetermined portion. In some embodiments, the cells grown on the substrate are separated into a plurality of randomly sized portions. Although substantially planar substrates are described and illustrated herein, substrates include non-planar substrates, such as curved substrates. For example, a flow of fluid through the substrate may apply a predetermined shear force to separate the meat product from the substrate.

[0197] FIG. 19 is a flow chart generally illustrating a method (1900) of preparing a meat product using any of the devices described herein. The process optionally includes rotating the device into a first configuration (1902). In some embodiments, the substrate may be rotated parallel to the ground. Cells are delivered to the surface of the substrate (1309). In some embodiments, at least about 500 cells / cm. 2 In some embodiments, about 5,000 cells / cm are provided on the substrate. 2 to approximately 1,000,000 pieces / cm 2 , including all values ​​and subranges therebetween, are delivered to the substrate until they attach to the surface of the substrate.

[0198] For example, the first configuration described above may be configured to seed a first side of one or more substrates with a plurality of cells such that the cells settle by gravity and adhere and / or attach to the first side of the substrate. For example, cells suspended in a fluid medium may be flowed over substantially the entire surface area of ​​the substrate. In the case of a flat substrate, the substrate may be positioned (e.g., rotated) to be parallel to the ground. In some of these embodiments, the first configuration of the device may further enable seeding a second side (e.g., the opposite side) of one or more substrates with a plurality of cells, thereby seeding the cells over substantially the entire surface of the substrate.

[0199] In some embodiments, the substrate may be rolled up as shown in Figures 16A-16D. The rolled substrate may be placed in a cavity of the device. For example, the rolled substrate may be naturally unfolded when placed in the cavity and held in the space within the device.

[0200] In some embodiments, the substrate can be prepared for cell seeding by wetting the exterior surface of the substrate. For example, a coating of growth medium can be applied to the exterior surface of the substrate. The device can be selectively filled with growth medium and removed. Cell populations are then placed on each region of the substrate.

[0201] After seeding the substrates, the cells are cultured to grow a meat product. The apparatus may be rotated (1906) to a second configuration. For example, the second configuration may be configured to allow cellular fluid flow to one or more substrates. In some embodiments, the substrates may be aligned substantially perpendicular to the ground. For example, multiple substrates may be arranged in a parallel plate configuration with the substrates perpendicular to the ground, thereby allowing fluid to flow through the spaces between the substrates. The cells adhere to the substrates such that they remain on the substrates against gravity. In some embodiments, the substrates may be positioned at an acute angle with the ground.

[0202] A first fluid comprising growth medium may flow through the device (1908). In some embodiments, the system may be configured to continuously culture cells for a predetermined period of time, such as at least one day. In some embodiments, the first fluid within the device may be recirculated at a speed of up to about 3.0 meters per second. In some embodiments, the first fluid flows through the device from a first height to a second height that is lower than the first height. This configuration may facilitate complete circulation of the fluid within the device. In some embodiments, the first fluid forms a planar flow through the substrate to provide a non-tortuous flow path that further facilitates consistent cell growth and complete exchange of the first fluid throughout the substrate.

[0203] In some embodiments, the fluid may be distributed to the substrate in a predetermined flow pattern, for example, the predetermined flow pattern may include a substantially uniform or laminar flow across a diameter of the substrate.

[0204] The cells on the substrate are cultured with the first fluid (1910) to produce a meat product. Thus, an edible meat product may be grown on the substrate. In some embodiments, culturing the cells includes providing a substantially uniform and stratified first fluid to the substrate, thereby providing consistent optimized cell growth across the entire surface of the substrate. The first fluid may include different compositions for different growth stages of the meat product. In some embodiments, the first fluid may flow generally downward through the substrate. Additionally or alternatively, the first fluid may flow generally upward through the substrate.

[0205] In some embodiments, the meat product may be held together by an extracellular matrix secreted by cells and / or cell-cell interactions. In some embodiments, the cells include one or more of myoblasts, mesangioblasts, myofibroblasts, mesenchymal stem cells, hepatocytes, fibroblasts, pericytes, adipocytes, epithelial, chondrocytes, osteoblasts, osteoclasts, pluripotent cells, somatic stem cells, and endothelial cells. In some embodiments, the cells include one or more cells from livestock, poultry, game, and aquatic animal species.

[0206] The meat product may have a thickness (e.g., height) of at least about 1 μm. For example, the meat product has a thickness in the range of about 1 μm to about 5 mm, about 100 μm to about 1 mm, about 200 μm to about 500 μm, about 300 μm to about 800 μm, including all values ​​and subranges therebetween.

[0207] One or more of the device, cells, and fluid may be measured (1912) to monitor the growth of the meat product. For example, one or more sensors may be configured to monitor one or more of the fluid, cell sheet, device, fluid pump, and fluid source. In some embodiments, the sensor measurements may be used to modify one or more fluid parameters, such as fluid flow rate and fluid composition. Additionally, the sensor measurements may be output to a user to allow the process to be monitored. Sensor measurements include, but are not limited to, pH, dissolved gas concentration, osmolality, nutrient concentration, waste concentration, ion concentration, oxygen concentration, temperature, and the like. In some embodiments, the device for preparing a meat product may include a temperature range of about 4° C. to about 40° C., a pH of about 6.6 to about 7.8, a dissolved oxygen of about 2% to about 120%, and all values ​​and subranges therebetween.

[0208] Once the growth of the meat product reaches a predetermined criterion, the meat product may be separated from the substrate for collection. In some embodiments, a second fluid may be flowed into the apparatus (1914). The meat product may be separated from the substrate utilizing the second fluid (1916). When sufficient shear force is applied to the meat product by the second fluid having a predetermined rate, the meat product separates from the substrate. In some embodiments, the second fluid may include a non-laminar turbulent flow. For example, the second fluid may include a gas and liquid (e.g., gas bubbles in a liquid) configured to generate a turbulent fluid and / or a periodic fluid flow pattern to aid in the separation of the meat product from the substrate. In some embodiments, the second fluid may include a saline solution (e.g., 0.2-0.9% saline). In some embodiments, the second fluid may be regulated by a second fluid diffuser (e.g., a spray ball) configured to separate the meat product from one or more substrates. In some variations, the edible meat product may be separated from the substrate by spraying the fluid at a predetermined angle relative to the substrate. The predetermined angle may be between about 10 degrees and approximately parallel to the substrate. In some embodiments, the separator may be configured to output fluid from one or more fluid nozzles to spray the fluid onto the substrate at a predetermined angle. In some embodiments, the separator may be configured to rotate about a predetermined axis when spraying the fluid onto the meat products. The fluid may have a linear velocity between about 0.003 m / sec and about 3.0 m / sec. The fluid may have a flow path having an angle of up to about 120 degrees.

[0209] In some embodiments, the meat product may be separated as a substantially continuous and / or whole multi-layer piece. That is, the substrate and second fluid allow the meat product to be peeled from the substrate without breaking and / or simultaneously. The meat product need not be separated as a single piece, but may be separated in predetermined portions rather than breaking apart in any uncontrolled manner. In some embodiments, the meat product may be separated in multiple separable portions (e.g., two or more portions).

[0210] In some embodiments, separation of the meat product from the substrate may be performed using multiple mechanisms. For example, separation of the meat product from the substrate may include one or more of fluidic, spontaneous, chemical, electrical, optical, thermal, and mechanical separation by inducing contraction and / or relaxation of the cells. For example, one or more buffers or enzyme solutions contact the cells to induce detachment from the substrate. In some embodiments, at least one of the volume and velocity of fluid flow through the fluidic channel may be increased to separate the meat product. For example, the fluid flow may be increased between adjacent substrates in a parallel plate configuration to fluidically separate the meat product from the substrate. In some variations, the fluid may be removed from the device prior to separating the edible meat product from the substrate. As another example, a second fluid containing air bubbles is flowed over the surface of the substrate to create a turbulent flow configured to apply sufficient shear force to separate the cell sheet from the substrate. In some embodiments, the second fluid has a higher viscosity than the first fluid, which may increase the interfacial shear stress applied to the cell sheet, thereby facilitating detachment of the cell sheet. In some embodiments, the second fluid may have the same composition as the first fluid.

[0211] The meat product is separated from the substrate and may be collected within or outside of the apparatus. In some embodiments, the separated meat product may be collected (1918). In some embodiments, substantially the entire meat product may be released (e.g., controlled, shortened, extended) at a predetermined time, whereby substantially only the cell sheet remains attached to the substrate or breaks down. Additionally or alternatively, the meat product may be separated into separable portions. For example, the substrate may be configured to allow the meat product grown on the substrate to be separated into multiple portions (e.g., two or more portions). The meat product may be collected within or outside of the apparatus for preparing the meat product. In some embodiments, the meat product may be separated into portions having a size of about 0.0001 m. 3 About 0.1m from 3 The volume may be between 0.01 and 0.1.

[0212] The device may be sterilized for reuse (1920) to at least reduce waste and costs. For example, the device may be steam sterilized before culturing and separating another meat product on the substrate. In some embodiments, the device may be disassembled. For example, at least one of the substrate, fluid diffuser, and collector may be removed from the device. In some embodiments, the substrate, fluid diffuser, and collector include flat, thin surfaces configured to drain freely and have no joints or internal surfaces (e.g., internal gaps, spaces) that may trap debris. Linear fluid channels also aid in sterilization.

[0213] IV. example As described herein, the systems, devices, and methods provided herein are useful for controlling the growth, attachment, release, and / or retention characteristics of cultured cells (e.g., cell sheets) for preparing meat products. Table 1 (below) lists the amount of meat product produced with different cell species and cell types over different time periods using the systems and devices described herein.

[0214] Table 1 TIFF0007633447000001.tif60135As used herein, the terms "about" and / or "approximately" when used in conjunction with a numerical value and / or range generally refer to a numerical value and / or range that is close to the stated numerical value and / or range. In some cases, the terms "about" and "approximately" may mean within ±10% of the stated value. For example, in some cases, "about 100 [units]" may mean within ±10% of 100 (e.g., 90 to 110). The terms "about" and "approximately" may be used interchangeably.

[0215] Although the foregoing implementations have been described in some detail by way of illustration and example for purposes of clarity and understanding, it will be apparent that certain changes and modifications may be made and are intended to be within the scope of the appended claims. It will be further understood that the components and features of the substrates described herein may be used in any combination, and the methods described herein may include all or a portion of the elements described herein. The description of a particular element or feature with respect to a particular figure is not intended to be limiting, and should not be construed to suggest that the element cannot be used in combination with other described elements.

[0216] Moreover, any combination of two or more such features, structures, systems, articles, materials, kits, steps, and / or methods disclosed herein is included within the inventive scope of the present disclosure, provided such features, structures, systems, articles, materials, kits, steps, and / or methods are not mutually inconsistent. Furthermore, some embodiments disclosed herein may be distinguished over the prior art by specifically lacking one or more features, elements, and functionality found in a reference or combination of references (i.e., claims directed to such embodiments may include a negative limitation).

[0217] All references presented in this application to publications or other documents, including but not limited to patents, patent applications, articles, web pages, books, etc., are incorporated herein by reference in their entirety. Additionally, all definitions defined and used herein are to be understood beyond dictionary definitions, definitions within documents incorporated by reference, and / or the ordinary meaning of the defined terms.

Claims

1. providing an enclosure having a cavity and a substrate, the substrate being inclined relative to a ground surface such that a fluid medium flows in one direction along the substrate; flowing the cells suspended in the fluid medium across the substrate to form the cell-based meat.

20. A method for forming cell-based meat comprising:

2. 10. The method of claim 1, further comprising recirculating the cells suspended in the fluid medium over the substrate.

3. 2. The method of claim 1, wherein the substrate has two sides and cells adhere to both sides.

4. The method of claim 3, further comprising rotating the substrate while flowing the cells suspended in the fluid medium across the substrate.

5. The method of claim 1 , further comprising circulating one or more gases between the enclosures.

6. 6. The method of claim 5, wherein circulating the one or more gases between the enclosures comprises circulating oxygen between the enclosures.

7. 10. The method of claim 1, further comprising harvesting and transferring the cell-based meat to a second enclosure from which the fluid medium has been at least partially evacuated.

8. providing a housing having a cavity and a substrate, the substrate being planar in shape and tilted relative to a longitudinal axis of the housing such that liquid flows in one direction along the substrate; circulating a fluid containing cells across the substrate; Cultivating the cells into cell-based meat 16. A method for growing a cell-based meat product comprising:

9. 10. The method of claim 8, further comprising recirculating unattached cells across the substrate.

10. 9. The method of claim 8, wherein the substrate has two surfaces and circulates the fluid containing the cells.

11. 11. The method of claim 10, further comprising rotating the substrate while circulating a fluid containing the cells so as to seed both sides of the substrate with the one or more cells.

12. The method of claim 8 , further comprising circulating one or more gases between the enclosures.

13. 13. The method of claim 12, wherein circulating the one or more gases between the enclosures comprises circulating oxygen between the enclosures.

14. 10. The method of claim 8, further comprising harvesting and transferring the cell-based meat to a second enclosure from which the fluid has been at least partially evacuated.

15. providing an enclosure having a cavity and a substrate, the substrate being inclined relative to a ground surface such that a liquid flows in one direction along the substrate; circulating cells suspended in a nutrient rich medium over the substrate; contacting the substrate with the nutrient-rich medium at a first flow rate to circulate the nutrient-rich medium to cells adhered to the substrate; harvesting the cells adhered to the substrate from the substrate by contacting the cells adhered to the substrate with the nutrient-rich medium, a buffer solution, an enzyme solution, saline, a fluid containing air bubbles, a fluid having a higher viscosity than the nutrient-rich medium, or a combination thereof, at a second flow rate, the second flow rate being greater than the first flow rate; 16. A method for growing a cell-based meat product comprising:

16. 16. The method of claim 15, further comprising recirculating unattached cells across the substrate.

17. 16. The method of claim 15, wherein the substrate has two surfaces and the cells are circulated on both surfaces of the substrate.

18. 20. The method of claim 17, further comprising rotating the substrate while circulating the cells on both sides of the substrate.

19. The method of claim 15 further comprising circulating oxygen between the enclosures.

20. 16. The method of claim 15, wherein said harvesting further comprises transferring cells adherent to said substrate to a second housing from which said fluid has been at least partially evacuated.

21. The method of claim 15 , wherein the substrate has a planar shape.

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