Plant and animal cell blend meat product and method for producing the same

Abstract

[0003] Embodiments described herein relate to blended meat substitute products and methods of making the same. In some aspects, the method of making the meat substitute product can include providing a carrier material to a plant-based scaffold, the carrier material comprising animal cells, adding a gelling agent to at least one of the plant-based scaffolds, and incubating the carrier material and the plant-based scaffold in a controlled environment to produce the meat substitute product. In some embodiments, the animal cells can include at least one of skeletal muscle cells, adipocytes, connective tissue cells, or skin cells.

Description

[Technical Field]

[0001] Statement on Federally Sponsored Research This invention was made with government support under award 2112169 from the National Science Foundation. The government has certain rights in this invention.

[0002] (CROSS-REFERENCE TO RELATED APPLICATIONS) This application claims priority to U.S. Provisional Application No. 63 / 407,472, filed September 16, 2022, and entitled "Plant and Animal Cell Blended Meat Products and Methods of Producing the Same," the disclosure of which is incorporated herein by reference in its entirety.

[0003] FIELD OF THE INVENTION Embodiments described herein relate to plant and animal cell-based meat products and methods for producing the same. [Background technology]

[0004] Plant-based meats often have high nutritional value and minimal health drawbacks. For example, they can contain all essential amino acids. However, they do not contain the same proteins or fats found in animal tissues and often lack the sensory properties associated with meat derived from animals. Cell-based meat has recently emerged as a viable alternative to plant-based meat. Cell-based meat involves animal cells that are cultured and used to build muscle and / or fat tissue similar to that derived from animals. Scaling up cell-based meat is difficult and involves many obstacles. By combining the positive aspects of plant-based meat and animal cell-based meat, it is possible to obtain a product that has organoleptic properties similar to meat derived from live animals and can be manufactured at a competitive price point. Summary of the Invention

[0005] Embodiments described herein relate to blended meat substitute products and methods for producing the same. In some aspects, the method of producing the meat substitute product can include providing a carrier material to a plant-based scaffold, the carrier material comprising animal cells or recombinantly produced animal flavor-related compounds; adding a gelling agent to at least one of the plant-based scaffold or the carrier; and incubating the carrier material and the plant-based scaffold in a controlled environment to produce the meat substitute product. In some embodiments, the animal cells can comprise at least one of skeletal muscle cells, adipocytes, connective tissue cells, or skin cells. In some embodiments, the carrier can comprise a carrier liquid. In some embodiments, the carrier material can be a first carrier material, and the method can further include immersing the plant-based scaffold in a second carrier material, the second carrier material having a viscosity different from that of the first carrier material and a penetration depth into the plant-based scaffold different from that of the first carrier material. In some embodiments, the carrier material can harden within the scaffold by temperature treatment. In some embodiments, the carrier material can harden within the scaffold by ionic gelation. In some embodiments, cells can be cultured within the scaffold. In some embodiments, the penetration depth of the carrier material into the scaffold can be controlled by adjusting the water content of the scaffold. [Brief explanation of the drawings]

[0006] The patent or application file contains at least one drawing executed in color. Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0007] Optional items in all figures shown with dashed lines.

[0008] [Figure 1] FIG. 1 is a block diagram of a method for producing a plant and animal cell blended meat product according to one embodiment. [Figure 2]FIG. 1 is a block diagram of a blended meat product according to one embodiment. [Figure 3A] FIG. 1 is a diagram of a process for producing a blended meat product, according to one embodiment. [Figure 3B] FIG. 1 is a diagram of a process for producing a blended meat product, according to one embodiment. [Figure 3C] FIG. 1 is a diagram of a process for producing a blended meat product, according to one embodiment. [Figure 4A] Scaffold soaking with various levels of carrier liquid diffusion is shown. [Figure 4B] Scaffold soaking with various levels of carrier liquid diffusion is shown. [Figure 4C] Scaffold soaking with various levels of carrier liquid diffusion is shown. [Figure 4D] Scaffold soaking with various levels of carrier liquid diffusion is shown. [Figure 5A] FIG. 1 shows a detailed view of a plant-based meat substitute product with a transparent skin covering the scaffold. [Figure 5B] FIG. 1 shows a detailed view of a plant-based meat substitute product with a transparent skin covering the scaffold. [Figure 5C] FIG. 1 shows a detailed view of a plant-based meat substitute product with a transparent skin covering the scaffold. [Figure 6A] Scaffolds infiltrated by dipping and by injection are shown. [Figure 6B] Scaffolds infiltrated by dipping and by injection are shown. [Figure 7] An image of a plant-based meat alternative product. DETAILED DESCRIPTION OF THE INVENTION

[0009] The field of tissue engineering involves biocompatible materials used as scaffolds for the growth of biological cells and tissues. Cell types that form meat include myofibers in muscle and adipocytes in fat. Such cells are often anchorage-dependent and, as a result, depend on an attachment substrate for survival and proliferation. For this reason, a variety of attachment substrates have been developed over the past several decades. These attachment substrates range from small spherical supports used in suspension bioreactors to porous 3D matrices used in reaction beds. While tissue engineering applications are numerous, they are mostly limited to medical applications, with few aimed at the production of edible products.

[0010] Applications for the production of edible products generally fall into two categories: (1) traditional tissue engineering approaches, in which cells are cultured inside a scaffold, or (2) blended products, in which cells are grown using various culture methods and then combined with separate materials that add structure to the final product. Tissue engineering approaches often involve the long-term culture (i.e., days to weeks, sometimes months) of cells in a material scaffold, allowing the cells to mature and develop into dense tissue. Such methods are often performed on a small scale, with tissue thickness limited to less than 0.5 mm, because diffusion limits nutrient transport. Efforts to vascularize tissues are underway, but have not yet resulted in the production of thick tissues at the scale or cost relevant to food manufacturing. Viable tissue engineering applications in the near future include those focusing on thin tissues such as skin, cornea, or thin tissues coating medical devices to improve implant outcomes.

[0011] Blended product approaches have historically involved growing cells and then adding them to a support material. These methods approach price parity with meat derived from live animals, as the percentage of cells representing the final product is case-dependent and potentially low compared to most tissue engineering approaches. Cells are often collected in a high-density pellet and resuspended in a scaffold at a dilute density compared to the original cell pellet, animal meat tissue, or continuously cultured tissue-engineered products. This allows for significant cell expansion during culture. Blended products utilize edible base materials that can contain cells as additives to improve nutrition, aroma, and / or flavor. Blended products are manufactured using various methods to retain cells within their support material. However, when added to a support material simply as an aqueous solution, the distribution of cells within the material is difficult to control, and cells often simply flow through the material without being properly retained. Carrier liquids and gelling agents can help retain cells within the support material. Most natural tissues used in meat derived from live animals contain different arrangements of multiple cell types, which give rise to the characteristic properties of each meat. Blended products may also suffer from cellular immaturity because they are not given sufficient time to develop within a structural framework that directs their development along the pathways seen in native tissues. For this reason, long muscle fibers are unlikely to be present in current state-of-the-art blended products.

[0012] Fibrous plant-based scaffolds can replicate long muscle fiber morphology in terms of both length and diameter. Therefore, fibrous plant-based scaffolds can contribute texture (organoleptic properties) that are lacking when immature muscle cells are used. For this reason, blended products based on fibrous scaffolds can have better texture than 100% muscle tissue, as has been the case until now, when muscle tissue is immature compared to animal muscle tissue. 3D printing is another strategy used to impart "fibrous" texture, but it suffers from significant throughput limitations because material extrusion rate is inversely proportional to extrusion diameter, and muscle fiber diameters range from 10 micrometers to 150 micrometers, making 3D printing slow and impractical at that scale, even when using multiple nozzles.

[0013] The methods described herein relate to the combination of cultured cells with plant-based meat products to produce blended products containing both plant and animal cell components. The methods described herein facilitate the transfer of cells into and retention of cells within the plant-based material. Furthermore, the transfer of specific cell types to specific regions within the plant-based material can facilitate the production of layered or stratified tissue. Products resulting from the methods described herein can range from substantially plant-based, with few animal cells therein, to primarily animal cell-based. In some embodiments, cells can be grown within the plant-based material. The embodiments described herein can provide price parity with meat derived from live animals.

[0014] Some embodiments described herein may include the plant proteins and animal cells described in U.S. Provisional Patent Application No. 63 / 346,172, entitled "Plant-Based Shredded Meat Products, and Meat Products, and Methods of Producing the Same," filed May 26, 2022 (the "'172 Application"), the disclosure of which is incorporated herein by reference in its entirety.

[0015] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. Thus, for example, the term "element" is intended to mean a single element or a combination of elements, and "material" is intended to mean one or more materials or a combination thereof.

[0016] The term "substantially," when used in connection with "cylindrical," "linear," and / or other geometric relationships, is intended to convey that the structure so defined is nominally cylindrical, linear, etc. As an example, a portion of a support member described as "substantially linear" is intended to convey that while linearity of the portion is desired, some nonlinearity may occur in the "substantially linear" portion. Such nonlinearity may result from manufacturing tolerances or other practical considerations (e.g., pressure or force applied to the support member, etc.). Thus, a geometric configuration modified by the term "substantially" includes such geometric characteristics within a ±5% tolerance of the stated geometric configuration. For example, a "substantially linear" portion is one that defines an axis or centerline that is within ±5% of being linear.

[0017] As used herein, the terms "set" and "plurality" can refer to multiple features or a single feature with multiple portions. For example, when referring to a set of fibers, the set of fibers can be considered an electrode with multiple portions, or the set of electrodes can be considered multiple separate fibers. Thus, a set of portions or multiple portions can include multiple portions that are either continuous or discontinuous with one another. Multiple particles or multiple materials can also be fabricated from multiple items that are manufactured separately and later joined together (e.g., by mixing, adhesive, or any suitable method). The term "set" with respect to a carrier material can refer to the gelation or hardening of the carrier material, characterized by a sudden increase in viscosity or solidification. The carrier solution can harden within the scaffold by a change in temperature or the addition of a gelling agent.

[0018] As used herein, "plant" or "plant-based" can include any material used in food production that is not animal-based. In other words, "plant" or "plant-based" is not limited to organisms of the plant kingdom. For example, a "plant-based scaffold" as described herein should be understood to include fungal-derived products such as mycelium or plant-like protists such as seaweed or algae.

[0019] The term "progenitor cell" is used herein to refer to a cell that has a more primitive cellular phenotype (e.g., earlier along a developmental pathway or progression than a fully differentiated cell) and has a greater degree of potency compared to the cells that can result from differentiation. Often, progenitor cells also have significant or very high proliferative potential. Progenitor cells can give rise to multiple different differentiated cell types or a single differentiated cell type, depending on the developmental pathway and the environment in which the cell develops and differentiates.

[0020] As used herein, the term "stem cell" refers to an undifferentiated cell that can proliferate and give rise to many more progenitor cells that have the potential to generate a large number of mother cells that can then give rise to differentiated or differentiable daughter cells that either terminally differentiate or can further mature and / or differentiate. The daughter cells themselves can be induced to proliferate and produce progeny that subsequently differentiate into one or more mature cell types, while also retaining one or more cells of the parent's developmental potential. The term "stem cell" refers to a subset of progenitor cells that, under certain circumstances, have the ability or potential to differentiate into a more specialized or differentiated phenotype and, under certain circumstances, retain the ability to proliferate without substantial differentiation. In one embodiment, the term stem cell generally refers to a naturally occurring mother cell, whose progeny (progeny) often specialize in different directions through differentiation, e.g., by acquiring entirely individual characteristics, as occurs in the progressive diversification of embryonic cells and tissues. Cell differentiation is a complex process that typically occurs through many cell divisions. Differentiated cells may be derived from multipotent cells that are themselves derived from multipotent cells. Although each of these multipotent cells can be considered a stem cell, the range of cell types each can give rise to can vary considerably. Some differentiated cells also have the ability to give rise to cells of greater developmental potential. This ability may be natural or artificially induced by treatment with various factors. In many biological cases, stem cells are also "multipotent" because they can produce progeny of two or more different cell types, but this is not required for "stemness." Self-renewal is another classic part of the definition of stem cells. Theoretically, self-renewal can occur by either of two major mechanisms: stem cells can divide asymmetrically, with one daughter retaining a stem state and the other daughter expressing some distinct or other specific function and phenotype. Alternatively, some stem cells in a population can divide symmetrically into two stem cells, thus maintaining some stem cells in the population as a whole, while other cells in the population give rise only to differentiated progeny.Formally, cells that begin as stem cells can progress toward a differentiated phenotype, but can then "reverse" and re-express the stem cell phenotype, a term often referred to as "dedifferentiation" or "reprogramming" or "reverse differentiation."

[0021] The term "embryonic stem cells" refers to pluripotent stem cells of the inner cell mass of blastocysts (see U.S. Patent Nos. 5,843,780 and 6,200,806, the contents of which are incorporated herein by reference). Such cells can also be obtained from the inner cell mass of blastocysts derived from somatic cell nuclear transfer (see, for example, U.S. Patent Nos. 5,945,577, 5,994,619, and 6,235,970, the contents of which are incorporated herein by reference). The distinctive characteristics of embryonic stem cells define the embryonic stem cell phenotype. Thus, if a cell has one or more of the unique characteristics of embryonic stem cells that allow it to be distinguished from other cells, the cell has the embryonic stem cell phenotype. Exemplary characteristic embryonic stem cell characteristics include, but are not limited to, gene expression profile, proliferation potential, differentiation potential, karyotype, responsiveness to specific culture conditions, etc.

[0022] The terms "adult stem cell" or "ASC" are used to refer to any multipotent stem cell derived from non-embryonic tissues, including fetal, juvenile, and adult tissues. Stem cells have been isolated from a wide variety of adult tissues, including blood, bone marrow, brain, olfactory epithelium, skin, pancreas, skeletal muscle, and cardiac muscle. Each of these stem cells can be characterized based on gene expression, factor responsiveness, and morphology in culture. Exemplary adult stem cells include neural stem cells, neural crest stem cells, mesenchymal stem cells, hematopoietic stem cells, and pancreatic stem cells.

[0023] Figure 1 is a block diagram of a method 10 for producing a plant and animal cell blended meat product according to one embodiment. As shown, method 10 includes providing a plant-based scaffold in step 11. Method 10 optionally includes seeding the plant-based scaffold with animal cells in step 12 and immersing the plant-based scaffold in a liquid in step 13. Method 10 further includes adding a first carrier material containing animal cells to the plant-based scaffold in step 14 and gelling the first carrier material within the plant-based scaffold in step 15. Method 10 optionally includes adding a second carrier material containing animal cells to the plant-based scaffold in step 16, the second carrier material containing animal cells. Then, in step 17, the carrier material and plant-based scaffold are incubated in a controlled environment to produce the meat substitute product.

[0024] Step 11 involves providing a plant-based scaffold. In some embodiments, the plant-based scaffold can comprise plant fibers. In some embodiments, the plant fibers can comprise bast fibers, leaf fibers, plant polysaccharides, starch, beta-glucan, cellulose, pectin polysaccharides, and / or seed hair fibers. In some embodiments, the plant fibers can include fibers derived from flax, hemp, Indian hemp, jute, tsinga, jute, kenaf, ramie, roselle, sunhemp, Bontenka, Manila hemp, kantara, henequen, maguey, Mauritus hemp, maolan, sisal, akund floss, bagasse, bamboo, bombax cotton, coir, cotton, floss silk tree, kapok, milkweed floss, or any combination thereof. In some embodiments, the plant-based scaffold can comprise plant proteins. In some embodiments, the plant protein can include protein derived from rice, pea, soybean, barley rice, bean, fava bean, seitan, tempeh, edamame, lentil, chickpea, nutritional yeast, spelt, teff, seeds, hemp seed, amaranth, quinoa, spirulina, green pea, oat, Ezekiel bread, wild rice, nuts, chia seed, mycoprotein, mycelium, or any combination thereof. In some embodiments, the plant protein can include one or more amino acids. In some embodiments, the plant protein can include alanine, arginine, asparagine, aspartic acid, cysteine, glutamine, glutamic acid, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, valine, or any combination thereof. In some embodiments, the plant-based scaffold can comprise an oil derived from a plant. In some embodiments, the oil can be food safe. In some embodiments, the oil can be organic.In some embodiments, the oil can comprise palm oil, canola oil, flaxseed oil, sunflower oil, soybean oil, corn oil, cottonseed oil, olive oil, coconut oil, peanut oil, safflower oil, sesame oil, almond oil, beech nut oil, Brazil nut oil, cashew oil, hazelnut oil, macadamia oil, mongongo nut oil, pecan oil, pine nut oil, pistachio oil, walnut oil, pumpkin seed oil, or any combination thereof.

[0025] In some embodiments, the plant-based scaffold can be grown or generated in a laboratory. In some embodiments, the plant-based scaffold can be pre-fabricated (e.g., purchased from a supplier). In some embodiments, the plant-based scaffold can be provided in its natural shape. In some embodiments, the plant-based scaffold can be formed into a desired shape to facilitate diffusion of carrier materials into the plant-based scaffold. In some embodiments, the plant-based scaffold can be molded. In some embodiments, the plant-based scaffold can be 3D printed. In some embodiments, the plant-based scaffold can have a tubular shape, a substantially linear shape, a spherical shape, or any other suitable shape factor, or a combination thereof. Further examples of scaffold shape factors are described in U.S. Patent Publication No. 2020 / 0330644, filed October 16, 2018, entitled "Methods of Forming Three-Dimensional Tissues Scaffolds Using Biological Fiber Inks and Methods of Use Thereof," the disclosure of which is incorporated herein by reference in its entirety.

[0026] Step 12 is optional and includes seeding the plant-based scaffold with animal cells. In some embodiments, the animal cells can be seeded onto a filamentous fungus-derived product. In some embodiments, the filamentous fungus-derived product can include mycelium. In some embodiments, the plant-based scaffold can be prefabricated with animal cells therein. In some embodiments, step 12 can include loading the plant-based scaffold with muscle cells, blending with fat cells, and optionally covering with skin cells. In some embodiments, step 12 can include delivering muscle and fat cells deep within the plant-based scaffold. In some embodiments, the animal cells can be seeded onto the plant-based scaffold by injection. In some embodiments, the animal cells can be seeded onto the plant-based scaffold by immersion. In some embodiments, a carrier containing animal cells can be added to the top surface of the scaffold and the scaffold can be immersed and / or infiltrated. In some embodiments, the animal cells can include skeletal muscle cells, fat cells, connective tissue cells, skin cells, or any combination thereof. In some embodiments, the scaffold may include mammalian cells, fish cells, avian muscle myoblasts, myoblasts from mammals, myoblasts from birds, myoblasts from fish, myosatellites, fibroblasts, adipocytes, endothelial cells, epithelial cells, keratinocytes, stem cells, or any combination thereof. In some embodiments, each cell type may be combined with a specific carrier solution with properties tailored to deliver the cells to specific regions within the scaffold.

[0027] Step 13 is optional and includes soaking the plant-based scaffold in a liquid. In some embodiments, soaking the plant-based scaffold can promote infiltration of a carrier material (e.g., a carrier liquid) into the plant-based scaffold. In some embodiments, soaking the plant-based scaffold in a liquid can promote gelation of the carrier material. In some embodiments, the liquid used to soak the plant-based scaffold can include salt to manipulate the osmotic pressure in the plant-based scaffold and open pores in the plant-based scaffold. In some embodiments, the liquid used to soak the plant-based scaffold can include a liquid that is free or substantially free of salt, such that the liquid manipulates the osmotic pressure in the plant-based scaffold and draws salt from the plant-based scaffold. Drawing salt from the plant-based scaffold can promote opening of pores in the plant-based scaffold. In some embodiments, the liquid used to soak the plant-based scaffold can include water, ethanol, glycerol, or any combination thereof. In some embodiments, the liquid used to soak the plant-based scaffold can aid in ionic gelation of the polysaccharide carrier. In some embodiments, the liquid used to soak the plant-based scaffold can include calcium lactate, calcium chloride, magnesium lactate, a potassium-containing compound (e.g., potassium chloride), and / or magnesium chloride. In some embodiments, the liquid used to soak the plant-based scaffold can aid in thermal gelation. The hydration state of the plant-based scaffold can be a factor in the plant-based scaffold's ability to absorb the carrier material and gelling agent. In some embodiments, the liquid used to soak the plant-based scaffold can be maintained at a temperature that induces gelation of the first carrier material. In some embodiments, the liquid used to soak the plant-based scaffold can include hydrocolloid, methylcellulose, kappa carrageenan, iota carrageenan, or any combination thereof. In some embodiments, the liquid used to soak the plant-based scaffold can be food-safe. In some embodiments, step 13 can include draining the liquid used to soak the plant-based scaffold.

[0028] In some embodiments, step 13 can result in a plant-based scaffold having a moisture content (i.e., before immersing the plant-based scaffold in the carrier material) of at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, or at least about 90% by weight. In some embodiments, step 13 can result in a plant-based scaffold having a moisture content of about 95% or less, about 90% or less, about 85% or less, about 80% or less, about 75% or less, about 70% or less, about 65% or less, about 60% or less, about 55% or less, about 50% or less, about 45% or less, or about 40% or less by weight. Combinations of the above moisture contents are also possible (e.g., at least about 35% and not more than about 95% by weight, or at least about 40% and not more than about 90% by weight), including all values ​​and ranges therebetween. In some embodiments, step 13 can result in a plant-based scaffold having a moisture content of about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95% by weight.

[0029] Step 14 comprises adding a first carrier material to the plant-based scaffold. The first carrier material comprises animal cells. In some embodiments, the first carrier material can comprise a carrier liquid. In some embodiments, the carrier liquid can be infused into the plant-based scaffold. In some embodiments, the carrier liquid can be heated. In some embodiments, the carrier liquid can undergo gelation upon heating. In some embodiments, the first carrier material can comprise a carrier gas. In some embodiments, the first carrier material can comprise a gel. In some embodiments, the animal cells can be mixed with the first carrier material before adding the first carrier material to the plant-based scaffold. In some embodiments, the animal cells can comprise skeletal muscle cells, adipocytes, connective tissue cells, skin cells, or any combination thereof. In some embodiments, the animal cells can comprise mammalian cells, fish cells, avian muscle myoblasts, fibroblasts, adipocytes, endothelial cells, epithelial cells, keratinocytes, stem cells, or any combination thereof. In some embodiments, the animal cells in the first carrier material can have a concentration of at least about 50,000 cells / ml, at least about 100,000 cells / ml, at least about 500,000 cells / ml, at least about 1,000,000 cells / ml, at least about 5,000,000 cells / ml, at least about 10,000,000 cells / ml, or at least about 50,000,000 cells / ml. In some embodiments, the animal cells in the first carrier material can have a concentration of about 100,000,000 cells / ml or less, about 50,000,000 cells / ml or less, about 10,000,000 cells / ml or less, about 5,000,000 cells / ml or less, 1,000,000 cells / ml or less, about 500,000 cells / ml or less, about 100,000 cells / ml or less, or about 50,000 cells / ml or less. Combinations of the above cell concentrations (e.g., at least about 50,000 cells / ml and about 100,000,000 cells / ml or less, or at least about 100,000 cells / ml and about 1,000,000 cells / ml or less) are also possible, including all values ​​and ranges therebetween.In some embodiments, the animal cells in the first carrier material can have a concentration of about 50,000 cells / ml, about 100,000 cells / ml, about 500,000 cells / ml, about 1,000,000 cells / ml, about 5,000,000 cells / ml, about 10,000,000 cells / ml, about 50,000,000 cells / ml, or about 100,000,000 cells / ml.

[0030] In some embodiments, the first carrier material can be water-based, alcohol-based, or oil-based. The carrier material can determine the extent to which animal cells penetrate the scaffold. In some embodiments, the first carrier material can undergo ionic gelation. In some embodiments, the first carrier material can include a polysaccharide such as pectin, chitosan, alginate, or any combination thereof. Dilute solutions of carrier liquids that undergo ionic gelation can be used for deep infiltration of muscle cells into plant-based scaffolds, while concentrated solutions can be used to create skin. In some embodiments, the first carrier material can include agar, pectin, alginate, carrageenan, gellan, gelatin, modified starch, methylcellulose, hydroxypropylmethylcellulose, or any combination thereof. In some embodiments, the first carrier material can include a polysaccharide dissolved in water. In some embodiments, the amount of polysaccharide dissolved in water can depend on the desired viscosity of the first carrier material.

[0031] In some embodiments, the first carrier material can undergo thermal gelation. In some embodiments, the first carrier material can include starch, modified starch, methylcellulose, polysaccharides, and / or plant proteins. In some embodiments, starch can be added to the scaffold before adding methylcellulose. In some embodiments, curdlan and / or konjac glucomannan can be added to the scaffold as a thermosetting polymer. In some embodiments, the first carrier material can be aqueous. In some embodiments, the first carrier material can be edible. In some embodiments, the first carrier material can be food-safe. In some embodiments, the first carrier material can be mixed with a fat substitute. In some embodiments, the first carrier material can include water and ethanol. In some embodiments, the first carrier material can include water containing monovalent ions, divalent ions, salts of monovalent ions, salts of divalent ions, or any combination thereof. In some embodiments, the monovalent ions and / or divalent ions can be cations, anions, or mixtures thereof. In some embodiments, the salt can include protons (or hydronium ions) or hydroxide ions to adjust the pH. In some embodiments, the first carrier material can be formulated to deliver cells to predetermined regions in the plant-based scaffold and to regions surrounding the plant-based scaffold. The carrier material can be formulated to retain the cells and prevent them from leaking out of the product. In some embodiments, the first carrier material can be formulated to render any fat contained in the first material, causing the product to sizzle when cooked.

[0032] In some embodiments, the first carrier material can be organic (i.e., associated with or derived from biological material). In some embodiments, the first carrier material can be certified organic as defined by the United States Department of Agriculture (USDA). In some embodiments, the first carrier material can be composed of ingredients produced through a process overseen by the USDA National Organic Program (NOP) and / or its certification agency. In some embodiments, the ingredients of the first carrier material can be produced in accordance with USDA regulations for certifying the organic character of ingredients. In some embodiments, the ingredients of the first carrier material can be produced using "certified materials" for organic certification, as specified by the USDA in 7 USC § 205(g). In some embodiments, the first carrier material can include ingredients that are 100% organic by weight, excluding salt and water, as defined by the USDA (i.e., the ingredients can meet the USDA's "100% organic" labeling criteria). In some embodiments, the first carrier material can include ingredients that are at least 95% organic by weight, excluding salt and water, as defined by the USDA (i.e., the ingredients can meet the criteria for the USDA's "organic" label). In some embodiments, the first carrier material can include ingredients that are at least 70% organic by weight, excluding salt and water, as defined by the USDA (i.e., the ingredients can meet the criteria for the USDA's "Made with Organic" label).

[0033] In some embodiments, the first carrier material can have a specific penetration depth into the plant-based scaffold. In some embodiments, the penetration depth of the first carrier material can be related to the viscosity of the first carrier material. In some embodiments, the viscosity of the first carrier material can be inversely proportional to the penetration depth of the first carrier material. In other words, the viscosity of the first carrier material can be controlled (e.g., by adding a specific amount of gelling agent) to control the penetration depth of the first carrier material. In some embodiments, animal cell types can be adapted to specific carrier liquids (e.g., muscle cells in a dilute carrier liquid that infiltrates the bulk scaffold volume, and skin cells in a concentrated carrier liquid that subsequently coats the surface of the plant-based scaffold). The penetration depth of the first carrier liquid can be a reliable metric for measuring the diffusion of a sample added to the surface of the plant-based scaffold. In some embodiments, the first carrier material can be added to the plant-based scaffold by dipping and / or soaking the plant-based scaffold in the first carrier material. In some embodiments, the first carrier material can be added to the plant-based scaffold by injecting the first carrier material into the plant-based scaffold. In some embodiments, the first carrier material and / or the second carrier material can be heat-set. In some embodiments, the first carrier material and / or the second carrier material can be gelled by exposure to a temperature difference. In some embodiments, the first carrier material and / or the second carrier material can be gelled when exposed to a temperature higher than the mixing temperature. For example, temperatures ranging from about 50°C to about 120°C can be used to gelatinize various starches.

[0034] In some embodiments, the first support material and / or the second support material can be heated to a temperature of at least about 30°C, at least about 40°C, at least about 50°C, at least about 60°C, at least about 70°C, at least about 80°C, at least about 90°C, at least about 100°C, at least about 100°C, at least about 110°C, at least about 120°C, at least about 130°C, at least about 140°C, at least about 150°C, at least about 160°C, at least about 170°C, at least about 180°C, at least about 190°C, at least about 200°C, at least about 210°C, at least about 220°C, at least about 230°C, at least about 240°C, at least about 250°C, at least about 260°C, at least about 270°C, at least about 280°C, or at least about 290°C. In some embodiments, the first support material and / or the second support material can be heated to a temperature of about 300°C or less, about 290°C or less, about 280°C or less, about 270°C or less, about 260°C or less, about 250°C or less, about 240°C or less, about 230°C or less, about 220°C or less, about 210°C or less, about 200°C or less, about 190°C or less, about 180°C or less, about 170°C or less, about 160°C or less, about 150°C or less, about 140°C or less, about 130°C or less, about 120°C or less, about 110°C or less, about 100°C or less, about 90°C or less, about 80°C or less, about 70°C or less, about 60°C or less, about 50°C or less, or about 40°C or less. Combinations of the above temperatures are also possible (e.g., at least about 30°C and not more than about 300°C, or at least about 50°C and not more than about 140°C), including all values ​​and ranges therebetween. In some embodiments, the first support material and / or the second support material can be heated to a temperature of about 30°C, about 40°C, about 50°C, about 60°C, about 70°C, about 80°C, about 90°C, about 100°C, about 100°C, about 110°C, about 120°C, about 130°C, about 140°C, about 150°C, about 160°C, about 170°C, about 180°C, about 190°C, about 200°C, about 210°C, about 220°C, about 230°C, about 240°C, about 250°C, about 260°C, about 270°C, about 280°C, about 290°C, or about 300°C.

[0035] In some embodiments, gelation can be induced by injecting a first carrier material and then using heat. In some embodiments, to improve the uniformity of the distribution of the hydrogel resulting from heating the carrier solution, the scaffold can be compressed and / or massaged before heating. In some embodiments, the scaffold can be compressed during heating. In some embodiments, the first carrier material and / or the second carrier material can be injected and evenly distributed throughout the scaffold. In some embodiments, the first carrier material and / or the second carrier material can be injected into a localized region of the scaffold.

[0036] In some embodiments, animal cells can attach to the scaffold via an attachment substrate. In other words, the animal cells can be cultured under conditions that promote cell attachment. The attachment substrate can substantially reduce the time required for animal cells to attach to the scaffold. In some embodiments, the attachment substrate can comprise small spherical carriers used in suspension bioreactors (e.g., spherical carriers having a particle size of about 1 μm, about 2 μm, about 3 μm, about 4 μm, about 5 μm, about 6 μm, about 7 μm, about 8 μm, about 9 μm, about 10 μm, about 20 μm, about 30 μm, about 40 μm, about 50 μm, about 60 μm, about 70 μm, about 80 μm, about 90 μm, or about 100 μm, including all values ​​and ranges therebetween) and / or porous 3D matrices used in reaction beds. In some embodiments, the attachment substrate can be composed of a hydrogel. In some embodiments, the attachment substrate can be composed of the same material as the plant-based scaffold (e.g., a combination of polysaccharides and plant proteins).

[0037] Step 15 involves gelling the first carrier material within the plant-based scaffold. In some embodiments, the gelling can be induced by a gelling agent. In some embodiments, the gelling can be induced by ionic gelation. In some embodiments, the gelling can be induced by heat treatment. In some embodiments, the gelling agent can be added to the plant-based scaffold by injection. When the gelling agent is injected, the diffusion distance from the addition point of the coverage within the sample is measured. In some embodiments, the gelling agent can be mixed with the first carrier material before immersing the plant-based scaffold in the first carrier material. In some embodiments, the gelling agent can be mixed with the first carrier material after immersing the plant-based scaffold in the first carrier material. In some embodiments, the gelling agent can be plant-based. In some embodiments, the gelling agent can include hydrocolloid, methylcellulose, high-viscosity methylcellulose, methylcellulose E / F / K, kappa-carrageenan, iota-carrageenan, or any combination thereof. In some embodiments, the gelling agent can induce ionic gelation in the first carrier material. In some embodiments, the first support material is capable of undergoing gelation by ion exchange. In some embodiments, the first support material is capable of undergoing gelation by sodium-calcium exchange.

[0038] In some embodiments, gelation can be by temperature treatment (e.g., heat treatment). In some embodiments, the first carrier material in the plant-based scaffold can be temperature treated to a temperature of at least about 60°C, at least about 65°C, at least about 70°C, at least about 75°C, at least about 80°C, at least about 85°C, at least about 90°C, at least about 95°C, at least about 100°C, at least about 105°C, at least about 110°C, or at least about 115°C. In some embodiments, the first carrier material in the plant-based scaffold can be temperature treated to a temperature of about 120°C or less, about 115°C or less, about 110°C or less, about 105°C or less, about 100°C or less, about 95°C or less, about 90°C or less, about 85°C or less, about 80°C or less, about 75°C or less, about 70°C or less, or about 65°C or less. Combinations of the above temperatures are also possible (e.g., at least about 60°C and not more than about 120°C, or at least about 80°C and not more than about 100°C), including all values ​​and ranges therebetween. In some embodiments, the first carrier material in the plant-based scaffold can be temperature treated to a temperature of about 60°C, about 65°C, about 70°C, about 75°C, about 80°C, about 85°C, about 90°C, about 95°C, about 100°C, about 105°C, about 110°C, about 115°C, or about 120°C. In some embodiments, gelling can be by steam treatment.

[0039] In some embodiments, the plant-based scaffold can be soaked in a gelling agent before adding the animal cells to the carrier material (i.e., before step 14). In other words, the plant-based scaffold can be pre-soaked in a gelling agent. Pre-soaking can be performed such that the first carrier material gels upon contact with the gelling agent present in the scaffold. For example, pre-soaking the plant-based scaffold in calcium lactate or calcium chloride will gel when a pectin-based carrier is added to the plant-based scaffold. For such gelation to occur, the concentration of the pre-soaked gelling agent should be relatively low (i.e., less than about 5 wt%, less than about 4 wt%, less than about 3 wt%, less than about 2 wt%, or less than about 1 wt% of the weight of the plant-based scaffold) to allow penetration into the first carrier material without premature gelation. In some embodiments, the gelling agent can be added to the plant-based scaffold both before and after adding the first carrier material and animal cells to the plant-based scaffold. In some embodiments, a first gelling agent can be added to the plant-based scaffold before the addition of the first carrier material, and a second gelling agent can be added to the plant-based scaffold after the addition of the first carrier material.

[0040] In some embodiments, the plant-based scaffold is not pre-soaked in a gelling agent. In such embodiments, a first carrier material and animal cells are added to the plant-based scaffold, and the plant-based scaffold (including the first carrier material and animal cells) is soaked in a gelling agent. In such cases, the gelling agent is at a higher concentration (i.e., at least about 5%, at least about 6%, at least about 7%, at least about 8%, at least about 9%, or at least about 10% by weight of the plant-based scaffold) than in pre-soaking to induce immediate or rapid gelation so that the first carrier material and animal cells are held in place before they can be washed away.

[0041] In some embodiments, the gelling agent can be added stepwise after each combination of carrier liquid and animal cells. For example, a dilute carrier liquid containing muscle cells can be added to a plant-based scaffold, followed by gelation by adding a gelling agent. A concentrated carrier liquid containing animal cells (skin) can then be added as a scaffold coating, and the resulting product can then be further gelled.

[0042] In some embodiments, the gelling agent can have a concentration in the first carrier material of at least about 0.1 wt%, at least about 0.2 wt%, at least about 0.3 wt%, at least about 0.4 wt%, at least about 0.5 wt%, at least about 0.6 wt%, at least about 0.7 wt%, at least about 0.8 wt%, at least about 0.9 wt%, at least about 1 wt%, at least about 2 wt%, at least about 3 wt%, at least about 4 wt%, at least about 5 wt%, at least about 6 wt%, at least about 7 wt%, at least about 8 wt%, or at least about 9 wt%. In some embodiments, the gelling agent can have a concentration in the first carrier material of about 10% by weight or less, about 9% by weight or less, about 8% by weight or less, about 7% by weight or less, about 6% by weight or less, about 5% by weight or less, about 4% by weight or less, about 3% by weight or less, about 2% by weight or less, about 1% by weight or less, about 0.9% by weight or less, about 0.8% by weight or less, about 0.7% by weight or less, about 0.6% by weight or less, about 0.5% by weight or less, about 0.4% by weight or less, about 0.3% by weight or less, or about 0.2% by weight or less. Combinations of the above concentrations of gelling agent in the first carrier material (e.g., at least about 0.1% by weight and about 10% by weight or less, or at least about 0.5% by weight and about 8% by weight or less) are also possible, including all values ​​and ranges therebetween. In some embodiments, the gelling agent can have a concentration in the first carrier material of about 0.1 wt%, about 0.2 wt%, about 0.3 wt%, about 0.4 wt%, about 0.5 wt%, about 0.6 wt%, about 0.7 wt%, about 0.8 wt%, about 0.9 wt%, about 1 wt%, about 2 wt%, about 3 wt%, about 4 wt%, about 5 wt%, about 6 wt%, about 7 wt%, about 8 wt%, about 9 wt%, or about 10 wt%.

[0043] In some embodiments, prior to the addition of the gelling agent, the first carrier material has a viscosity of at least about 1 mPa·s, at least about 2 mPa·s, at least about 3 mPa·s, at least about 4 mPa·s, at least about 5 mPa·s, at least about 6 mPa·s, at least about 7 mPa·s, at least about 8 mPa·s, at least about 9 mPa·s, at least about 10 mPa·s, at least about 20 mPa·s, at least about 30 mPa·s, at least about 40 mPa·s, at least about 50 mPa·s, at least about 60 mPa·s, at least about 70 mPa·s, at least about 80 mPa·s, at least about 90 mPa·s, at least about 100 mPa·s , at least about 200 mPa·s, at least about 300 mPa·s, at least about 400 mPa·s, at least about 500 mPa·s, at least about 600 mPa·s, at least about 700 mPa·s, at least about 800 mPa·s, at least about 900 mPa·s, at least about 1,000 mPa·s, at least about 2,000 mPa·s, at least about 3,000 mPa·s, at least about 4,000 mPa·s, at least about 5,000 mPa·s, at least about 6,000 mPa·s, at least about 7,000 mPa·s, at least about 8,000 mPa·s, or at least about 9,000 mPa·s.In some embodiments, prior to addition of the gelling agent, the first carrier material has a viscosity of about 10,000 mPa·s or less, about 9,000 mPa·s or less, about 8,000 mPa·s or less, about 7,000 mPa·s or less, about 6,000 mPa·s or less, about 5,000 mPa·s or less, about 4,000 mPa·s or less, about 3,000 mPa·s or less, about 2,000 mPa·s or less, about 1,000 mPa·s or less, about 900 mPa·s or less, about 800 mPa·s or less, about 700 mPa·s or less, about 600 mPa·s or less, about 500 mPa·s or less, about 400 mPa·s or less. The viscosity may be about 100 mPa·s or less, about 300 mPa·s or less, about 200 mPa·s or less, about 100 mPa·s or less, about 90 mPa·s or less, about 80 mPa·s or less, about 70 mPa·s or less, about 60 mPa·s or less, about 50 mPa·s or less, about 40 mPa·s or less, about 30 mPa·s or less, about 20 mPa·s or less, about 10 mPa·s or less, about 9 mPa·s or less, about 8 mPa·s or less, about 7 mPa·s or less, about 6 mPa·s or less, about 5 mPa·s or less, about 4 mPa·s or less, about 3 mPa·s or less, or about 2 mPa·s or less. Combinations of the above viscosities are also possible (e.g., at least about 1 mPa·s and about 10,000 mPa·s or less, or at least about 100 mPa·s and about 1,000 mPa·s or less), including all values ​​and ranges therebetween. In some embodiments, prior to addition of the gelling agent, the first carrier material has a viscosity of about 1 mPa·s, about 2 mPa·s, about 3 mPa·s, about 4 mPa·s, about 5 mPa·s, about 6 mPa·s, about 7 mPa·s, about 8 mPa·s, about 9 mPa·s, about 10 mPa·s, about 20 mPa·s, about 30 mPa·s, about 40 mPa·s, about 50 mPa·s, about 60 mPa·s, about 70 mPa·s, about 80 mPa·s, about 90 mPa·s, about 100 mPa·s, about 200 mPa·s, about The viscosity may be about 300 mPa·s, about 400 mPa·s, about 500 mPa·s, about 600 mPa·s, about 700 mPa·s, about 800 mPa·s, about 900 mPa·s, about 1,000 mPa·s, about 2,000 mPa·s, about 3,000 mPa·s, about 4,000 mPa·s, about 5,000 mPa·s, about 6,000 mPa·s, about 7,000 mPa·s, about 8,000 mPa·s, about 9,000 mPa·s, or about 10,000 mPa·s.

[0044] The viscosity of the first carrier material can be a function of the desired penetration depth into the plant-based scaffold. For example, the amount of polysaccharide added to water affects the resulting viscosity. After the addition of a gelling agent, the first carrier material becomes a carrier gel. The first carrier material can be exposed to a gelling agent (e.g., a divalent salt diluted in water), and the first carrier material solidifies at a rate that depends on the concentrations of the first carrier material and the gelling agent. In some embodiments, gelation can be rapid, such that the first carrier material can be considered solid after exposure to the gelling agent, thereby holding the carrier gel in place. In some embodiments, the kinetics of gel hardening can be advantageous in determining the extent of penetration of the first carrier material into the plant-based scaffold before complete gelation.

[0045] In some embodiments, after addition of the gelling agent, the carrier gel has a viscosity of at least about 10 mPa·s, at least about 20 mPa·s, at least about 30 mPa·s, at least about 40 mPa·s, at least about 50 mPa·s, at least about 60 mPa·s, at least about 70 mPa·s, at least about 80 mPa·s, at least about 90 mPa·s, at least about 100 mPa·s, at least about 200 mPa·s, at least about 300 mPa·s, at least about 400 mPa·s, at least about 500 mPa·s, at least about 600 mPa·s, at least about 700 mPa·s, at least about 800 mPa·s, at least about 900 mPa·s, at least about 1,000 mPa·s, The viscosity of the composition may be at least about 2,000 mPa·s, at least about 3,000 mPa·s, at least about 4,000 mPa·s, at least about 5,000 mPa·s, at least about 6,000 mPa·s, at least about 7,000 mPa·s, at least about 8,000 mPa·s, at least about 9,000 mPa·s, at least about 10,000 mPa·s, at least about 20,000 mPa·s, at least about 30,000 mPa·s, at least about 40,000 mPa·s, at least about 50,000 mPa·s, at least about 60,000 mPa·s, at least about 70,000 mPa·s, at least about 80,000 mPa·s, or at least about 90,000 mPa·s.In some embodiments, after addition of the gelling agent, the carrier gel has a viscosity of about 100,000 mPa·s or less, about 90,000 mPa·s or less, about 80,000 mPa·s or less, about 70,000 mPa·s or less, about 60,000 mPa·s or less, about 50,000 mPa·s or less, about 40,000 mPa·s or less, about 30,000 mPa·s or less, about 20,000 mPa·s or less, about 10,000 mPa·s or less, about 9,000 mPa·s or less, about 8,000 mPa·s or less, about 7,000 mPa·s or less, about 6,000 mPa·s or less, about 5,000 mPa·s or less, about 4,000 mPa·s or less The viscosity of the composition may be about 3,000 mPa·s or less, about 2,000 mPa·s or less, about 1,000 mPa·s or less, about 900 mPa·s or less, about 800 mPa·s or less, about 700 mPa·s or less, about 600 mPa·s or less, about 500 mPa·s or less, about 400 mPa·s or less, about 300 mPa·s or less, about 200 mPa·s or less, about 100 mPa·s or less, about 90 mPa·s or less, about 80 mPa·s or less, about 70 mPa·s or less, about 60 mPa·s or less, about 50 mPa·s or less, about 40 mPa·s or less, about 30 mPa·s or less, or about 20 mPa·s or less. Combinations of the above viscosities are also possible (eg, at least about 10 mPa·s and less than or equal to about 100,000 mPa·s, or at least about 1,000 mPa·s and less than or equal to about 10,000 mPa·s), including all values ​​and ranges therebetween.In some embodiments, after addition of the gelling agent, the carrier gel has a viscosity of about 10 mPa·s, about 20 mPa·s, about 30 mPa·s, about 40 mPa·s, about 50 mPa·s, about 60 mPa·s, about 70 mPa·s, about 80 mPa·s, about 90 mPa·s, about 100 mPa·s, about 200 mPa·s, about 300 mPa·s, about 400 mPa·s, about 500 mPa·s, about 600 mPa·s, about 700 mPa·s, about 800 mPa·s, about 900 mPa·s, about 1,000 mPa·s, about 2,000 mPa·s, about 3,000 mPa·s, or about 4,000 mPa·s. The composition may have a viscosity of about 100 mPa·s, about 4,000 mPa·s, about 5,000 mPa·s, about 6,000 mPa·s, about 7,000 mPa·s, about 8,000 mPa·s, about 9,000 mPa·s, about 10,000 mPa·s, about 20,000 mPa·s, about 30,000 mPa·s, about 40,000 mPa·s, about 50,000 mPa·s, about 60,000 mPa·s, about 70,000 mPa·s, about 80,000 mPa·s, about 90,000 mPa·s, or about 100,000 mPa·s.

[0046] Step 16 is optional and involves adding a second carrier material to the plant-based scaffold. In some embodiments, the second carrier material can comprise a carrier liquid. In some embodiments, the second carrier material can comprise a carrier gas. In some embodiments, the second carrier material can comprise a gel. The second carrier material can comprise animal cells. In some embodiments, the animal cells can be mixed with the second carrier material before adding the second carrier material to the plant-based scaffold. In some embodiments, the second carrier material can be substantially different from the first carrier material. For example, the first carrier material can be water-based and the second carrier material can be oil-based. In some embodiments, the second carrier material can be immiscible with the first carrier material. In some embodiments, the second carrier material can be used to create an outer layer or "skin" of the final product. In some embodiments, the second carrier material can comprise a fat and / or oil. In some embodiments, the second carrier material can have any of the properties described above with respect to the first carrier material (e.g., "certified organic"). In some embodiments, the second carrier material can include a gelling agent. In some embodiments, the gelling agent included in the second carrier material can be the same as the gelling agent included in the first carrier material. In some embodiments, the first carrier material can include a first gelling agent, and the second carrier material can include a second gelling agent, where the second gelling agent is different from the first gelling agent. In some embodiments, the second carrier material can include methylcellulose, microcrystalline cellulose, kappa-carrageenan, iota-carrageenan, plant protein, agar, pectin, alginate, carrageenan, xanthan gum, gelatin, modified starch, methylcellulose, hydroxypropyl methylcellulose, gellan gum, curdlan, nanoparticles, konjac glucomannan, or any combination thereof. In some embodiments, the second carrier material and the cells disposed therein can be designed to mimic fat. In some embodiments, the second carrier material can be added to the plant-based scaffold by soaking and / or immersing the plant-based scaffold in the first carrier material.In some embodiments, the first carrier material can be added to the plant-based scaffold by injecting the first carrier material into the plant-based scaffold.

[0047] In some embodiments, the second carrier material can have a viscosity higher than the viscosity of the first carrier material, such as at least about 5 mPa·s, at least about 6 mPa·s, at least about 7 mPa·s, at least about 8 mPa·s, at least about 9 mPa·s, at least about 10 mPa·s, at least about 20 mPa·s, at least about 30 mPa·s, at least about 40 mPa·s, at least about 50 mPa·s, at least about 60 mPa·s, at least about 70 mPa·s, at least about 80 mPa·s, at least about 90 mPa·s, at least about 100 mPa·s, at least about 200 mPa·s, at least about 300 mPa·s, at least about 400 mPa·s, at least about 500 mPa·s, or at least about It can have a viscosity of 600 mPa·s, at least about 700 mPa·s, at least about 800 mPa·s, at least about 900 mPa·s, at least about 1,000 mPa·s, at least about 2,000 mPa·s, at least about 3,000 mPa·s, at least about 4,000 mPa·s, at least about 5,000 mPa·s, at least about 6,000 mPa·s, at least about 7,000 mPa·s, at least about 8,000 mPa·s, at least about 9,000 mPa·s, at least about 10,000 mPa·s, at least about 20,000 mPa·s, at least about 30,000 mPa·s, or at least about 40,000 mPa·s.In some embodiments, the second support material has a viscosity of about 50,000 mPa·s or less, about 40,000 mPa·s or less, about 30,000 mPa·s or less, about 20,000 mPa·s or less, about 10,000 mPa·s or less, about 9,000 mPa·s or less, about 8,000 mPa·s or less, about 7,000 mPa·s or less, about 6,000 mPa·s or less, about 5,000 mPa·s or less, about 4,000 mPa·s or less, about 3,000 mPa·s or less, about 2,000 mPa·s or less, about 1,000 mPa·s or less, about 900 mPa·s or less, or about 800 mPa·s or less. The viscosity may be about 700 mPa·s or less, about 600 mPa·s or less, about 500 mPa·s or less, about 400 mPa·s or less, about 300 mPa·s or less, about 200 mPa·s or less, about 100 mPa·s or less, about 90 mPa·s or less, about 80 mPa·s or less, about 70 mPa·s or less, about 60 mPa·s or less, about 50 mPa·s or less, about 40 mPa·s or less, about 30 mPa·s or less, about 20 mPa·s or less, about 10 mPa·s or less, about 9 mPa·s or less, about 8 mPa·s or less, about 7 mPa·s or less, or about 6 mPa·s or less. Combinations of the above viscosities are also possible (eg, at least about 5 mPa·s and less than or equal to about 50,000 mPa·s, or at least about 100 mPa·s and less than or equal to about 10,000 mPa·s), including all values ​​and ranges therebetween.In some embodiments, after addition of the gelling agent, the first carrier material has a viscosity of about 5 mPa·s, about 6 mPa·s, about 7 mPa·s, about 8 mPa·s, about 9 mPa·s, about 10 mPa·s, about 20 mPa·s, about 30 mPa·s, about 40 mPa·s, about 50 mPa·s, about 60 mPa·s, about 70 mPa·s, about 80 mPa·s, about 90 mPa·s, about 100 mPa·s, about 200 mPa·s, about 300 mPa·s, about 400 mPa·s, about 500 mPa·s, about 600 mPa·s, about 700 mPa·s, about 80 mPa·s, about 90 mPa·s, about 100 mPa·s, about 200 mPa·s, about 300 mPa·s, about 400 mPa·s, about 500 mPa·s, about 600 mPa·s, about 700 mPa·s, about 800 mPa·s, about 90 ... mPa·s, about 800 mPa·s, about 900 mPa·s, about 1,000 mPa·s, about 2,000 mPa·s, about 3,000 mPa·s, about 4,000 mPa·s, about 5,000 mPa·s, about 6,000 mPa·s, about 7,000 mPa·s, about 8,000 mPa·s, about 9,000 mPa·s, about 10,000 mPa·s, about 20,000 mPa·s, about 30,000 mPa·s, about 40,000 mPa·s, or about 50,000 mPa·s.

[0048] In some embodiments, the first carrier material and the second carrier material can have different material compositions. In some embodiments, the first carrier material can be a water-based carrier liquid, and the second carrier material can be oil / fat-based. In some embodiments, the first carrier material can be oil / fat-based, and the second carrier material can be water-based. In some embodiments, the first carrier material and the second carrier material can have similar material compositions but at different concentrations. In some embodiments, the first carrier material can include water with a first polysaccharide concentration, while the second carrier material can include water with a second polysaccharide concentration, where the second polysaccharide composition is greater than the first polysaccharide composition. The different concentrations result in different viscosities and scaffold penetration kinetics. In some embodiments, method 10 can include soaking the plant-based scaffold in a third carrier material, a fourth carrier material, a fifth carrier material, a sixth carrier material, a seventh carrier material, an eighth carrier material, a ninth carrier material, or a tenth carrier material. Any combination of the aforementioned carrier materials is also possible. For example, a plant-based scaffold can be soaked in one fat / oil-based carrier and two different pectin-based carriers. In some embodiments, a first carrier material can be delivered by a first method, and a second carrier material can be delivered by a second method. In some embodiments, a first carrier material can be delivered by dipping and / or soaking the plant-based scaffold in the first carrier material, and a second carrier material can be delivered by injecting the second carrier material into the plant-based scaffold. In some embodiments, a first carrier material can be delivered by injecting the first carrier material into the plant-based scaffold, and a second carrier material can be delivered by dipping and / or soaking the plant-based scaffold in the second carrier material.

[0049] In some embodiments, the second carrier material can be added to the plant-based scaffold at least partially simultaneously with the first carrier material. In some embodiments, the second carrier material can be added to the plant-based scaffold before the first carrier material. In some embodiments, the second carrier material can be added to the plant-based scaffold after the first carrier material. In some embodiments, the second carrier material can have a penetration depth into the plant-based scaffold that is less than the penetration depth of the first carrier material. In some embodiments, the second carrier material can comprise animal cells. In some embodiments, the second carrier material can comprise skeletal muscle cells, adipocytes, connective tissue cells, or skin cells. In some embodiments, the second carrier material can comprise mammalian cells, fish cells, avian muscle myoblasts, fibroblasts, adipocytes, endothelial cells, epithelial cells, keratinocytes, stem cells, or any combination thereof. In some embodiments, the first carrier material can comprise a first type of cells and the second carrier material can comprise a second type of cells, where the second type of cells are different from the first type of cells. In some embodiments, the first carrier material and the second carrier material can contain the same type of cells. In some embodiments, the first carrier material can be mixed with muscle myoblasts and delivered deep within the plant-based scaffold, while the second carrier material can be mixed with dermal fibroblasts to form a concentrated skin near the outer surface of the blended meat product. In some embodiments, the first carrier material can form fat, the second carrier material can form muscle, and the third carrier material can form skin. In some embodiments, the first carrier material that forms fat can undergo temperature-dependent gelation. In some embodiments, the second carrier material that forms muscle can undergo deep delivery with low-concentration ionic gelation. In some embodiments, the third carrier material that forms skin can undergo high-concentration ionic gelation on the surface of the plant and animal cell blended meat product.In some embodiments, a first carrier material can include a first type of cells, a second carrier material can include a second type of cells, and a third type of cells can be injected into an interior region encompassed by the first type of cells and the second type of cells. In some embodiments, the injection of the third type of cells can occur before macerating the first type of cells and the second type of cells. In some embodiments, the injection of the third type of cells can occur after macerating the first type of cells and the second type of cells.

[0050] Step 17 is optional and involves incubating the carrier material and plant-based scaffold in a controlled environment to produce the meat substitute product. In some embodiments, an osmotic concentration gradient can be established within the plant-based scaffold, causing the carrier material to draw inward toward the center of the plant-based scaffold, reducing or eliminating the concentration gradient during incubation. For example, the plant-based scaffold can have a higher salt concentration than the carrier material, such that liquid from the carrier material migrates toward the center of the plant-based scaffold during incubation, equalizing the salt concentration throughout the product. In some embodiments, muscle, fat, and skin cells are separated in the meat substitute product so that they replicate the tissue structure found in a particular cut of meat.

[0051] The presence of animal cells in the carrier material and / or plant-based scaffold may limit the temperature, pH, and osmolality at which incubation can occur. Therefore, the incubation temperature and pH are set to minimize cell death during incubation. In some embodiments, the temperature during incubation can be at least about -20°C, at least about -15°C, at least about -10°C, at least about -5°C, at least about 0°C, at least about 5°C, at least about 10°C, at least about 15°C, at least about 20°C, at least about 21°C, at least about 22°C, at least about 23°C, at least about 24°C, at least about 25°C, at least about 26°C, at least about 27°C, at least about 28°C, at least about 29°C, at least about 30°C, at least about 31°C, at least about 32°C, at least about 33°C, at least about 34°C, at least about 35°C, at least about 36°C, at least about 37°C, at least about 38°C, or at least about 39°C. In some embodiments, the temperature during incubation can be about 40° C. or less, about 39° C. or less, about 38° C. or less, about 37° C. or less, about 36° C. or less, about 35° C. or less, about 34° C. or less, about 33° C. or less, about 32° C. or less, about 31° C. or less, about 30° C. or less, about 29° C. or less, about 28° C. or less, about 27° C. or less, about 26° C. or less, about 25° C. or less, about 24° C. or less, about 23° C. or less, about 22° C. or less, about 21° C. or less, about 20° C. or less, about 15° C. or less, about 10° C. or less, about 5° C. or less, about 0° C. or less, about −5° C. or less, about −10° C. or less, or about −15° C. or less. Combinations of the above incubation temperatures (e.g., at least about −20° C. and about 40° C. or less, or at least about 25° C. and about 35° C. or less) are also possible, including all values ​​and ranges therebetween. In some embodiments, the temperature during incubation can be about -20°C, about -15°C, about -10°C, about -5°C, about 0°C, about 5°C, about 10°C, about 15°C, about 20°C, about 21°C, about 22°C, about 23°C, about 24°C, about 25°C, about 26°C, about 27°C, about 28°C, about 29°C, about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, or about 40°C.

[0052] In some embodiments, the incubation pH can be at least about 6.5, at least about 6.6, at least about 6.7, at least about 6.8, at least about 6.9, at least about 7, at least about 7.1, at least about 7.2, at least about 7.3, at least about 7.4, at least about 7.5, at least about 7.6, at least about 7.7, at least about 7.8, or at least about 7.9. In some embodiments, the incubation pH can be about 8 or less, about 7.9 or less, about 7.8 or less, about 7.7 or less, about 7.6 or less, about 7.5 or less, about 7.4 or less, about 7.3 or less, about 7.2 or less, about 7.1 or less, about 7 or less, about 6.9 or less, about 6.8 or less, about 6.7 or less, about 6.6 or less, about 6.5 or less, about 6.4 or less, about 6.3 or less, about 6.2 or less, or about 6.1 or less. Combinations of the above pH values ​​are also possible (e.g., at least about 6.5 and not more than about 8, or at least about 7 and not more than about 7.5), including all values ​​and ranges therebetween. In some embodiments, the incubation pH can be about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, or about 8.

[0053] In some embodiments, the incubation can be for a period of at least about 30 minutes, at least about 1 hour, at least about 2 hours, at least about 3 hours, at least about 4 hours, at least about 5 hours, at least about 6 hours, at least about 7 hours, at least about 8 hours, at least about 9 hours, at least about 10 hours, at least about 12 hours, at least about 14 hours, at least about 16 hours, at least about 18 hours, at least about 20 hours, at least about 22 hours, at least about 1 day, at least about 2 days, at least about 3 days, at least about 4 days, at least about 5 days, at least about 10 days, at least about 15 days, at least about 20 days, or at least about 25 days. In some embodiments, the incubation can be for a period of about 30 days or less, about 25 days or less, about 20 days or less, about 15 days or less, about 10 days or less, about 5 days or less, about 4 days or less, about 3 days or less, about 2 days or less, about 1 day or less, about 22 hours or less, about 20 hours or less, about 18 hours or less, about 16 hours or less, about 14 hours or less, about 12 hours or less, about 10 hours or less, about 9 hours or less, about 8 hours or less, about 7 hours or less, about 6 hours or less, about 5 hours or less, about 4 hours or less, about 3 hours or less, about 2 hours or less, or about 1 hour or less.

[0054] In some embodiments, the blended meat product can be formed without an incubation period. In some embodiments, osmotic pressure can be the primary mechanism by which carrier materials are drawn to their desired location. Scaffold porosity, carrier concentration and viscosity, and infiltration methods (e.g., passive diffusion, injection) can be used to control carrier delivery into the plant-based scaffold. When cells are incorporated into the plant-based scaffold, there is a narrow range of osmotic pressures over which the cells can survive.

[0055] Combinations of the above incubation times are also possible (e.g., at least about 30 minutes and up to about 30 days, or at least about 2 hours and up to about 18 hours), including all values ​​and ranges therebetween. In some embodiments, the incubation can be for a period of about 30 minutes, about 1 hour, about 2 hours, about 3 hours, about 4 hours, about 5 hours, about 6 hours, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 12 hours, about 14 hours, about 16 hours, about 18 hours, about 20 hours, about 22 hours, about 1 day, about 2 days, about 3 days, about 4 days, about 5 days, about 10 days, about 15 days, about 20 days, about 25 days, or about 30 days.

[0056] In some embodiments, the flavoring agent can be added to the carrier material and / or the plant-based scaffold before the incubation period. In some embodiments, the flavoring agent can be added to the carrier material and the plant-based scaffold during the incubation period. In some embodiments, the flavoring agent can be added to the carrier material and the plant-based scaffold after the incubation period. In some embodiments, the flavoring agent can include a flavor enhancer. In some embodiments, the flavoring agent can include an aroma enhancer. In some embodiments, the flavoring agent can include one or more spices. In some embodiments, the flavoring agent is selected from the group consisting of table salt, black pepper, paprika, oregano, anise, celery seed, cassia, catnip, cardamom, caraway, burnet, brown mustard, borage, black pepper, mustard seed, cumin, bergamot, basil, bay leaf, asafoetida, anise, angelica, allspice, cayenne pepper, chervil, chicory, chili pepper, cinnamon, cilantro, cloves, coriander, costmary, curry, dill, fennel, and fenugreek. Blended meat products may include, for example, basil, filet, ginger, grains of paradise, holy basil, horehound, horseradish, hyssop, lavender, lemon balm, lemongrass, lemon verbena, licorice, lovage, mace, marjoram, nutmeg, oregano, paprika, parsley, peppermint, poppy seeds, rosemary, rue, saffron, sage, savory, sesame seeds, sorrel, star anise, spearmint, tarragon, thyme, turmeric, vanilla, wasabi, or any combination thereof. In some embodiments, a color enhancer may be added before, during, and / or after the incubation period. In some embodiments, animal flavor compounds may be produced via recombinant sources (e.g., recombinant myoglobin) and added to the blended meat product.

[0057] 2 is a block diagram of a blended meat product 100 according to one embodiment. As shown, the blended meat product 100 includes a plant-based scaffold 110 and a first gel layer 130. The blended meat product 100 optionally includes a second gel layer 150. In some embodiments, the plant-based scaffold 110 and the first gel layer 130 can be substantially mixed together such that they collectively form a single layer of material. The combination of the plant-based scaffold 110 and the first gel layer 130 is referred to herein as an "internal layer." In some embodiments, the internal layer can remain separate or partially separate. In some embodiments, the internal layer can form a homogenous or substantially homogenous single layer.

[0058] In some embodiments, the plant-based scaffold 110 may swell during the formation of the blended meat product 100. In some embodiments, the plant-based scaffold 110 may swell to about 1.5 times, about 2 times, about 2.5 times, about 3 times, about 3.5 times, about 4 times, about 4.5 times, about 5 times, about 5.5 times, about 6 times, about 6.5 times, about 7 times, about 7.5 times, about 8 times, about 8.5 times, about 9 times, about 9.5 times, or about 10 times its original size (including all values ​​and ranges therebetween) during the production of the blended meat product 100. In some embodiments, the plant-based scaffold 110 may become substantially softer during the production of the blended meat product. In some embodiments, the plant-based scaffold 110 may become firmer after gelation. In some embodiments, the first gel layer 130 may be encapsulated or substantially encapsulated in the plant-based scaffold 110 during the production of the blended meat product 100. In some embodiments, the penetration depth of the first gel layer 130 within the plant-based scaffold 110, and therefore the degree of intermixing between the first gel layer 130 and the plant-based scaffold 110, can be controlled by the carrier solution and gelling agent formulation.

[0059] In some embodiments, the inner layer can have a first composition and the second gel layer 150 can have a second composition, where the second composition is different from the first composition. In some embodiments, the inner layer can have a first texture and the second gel layer 150 can have a second texture, where the second texture is different from the first texture. In some embodiments, the second gel layer 150 can have a skin-like texture. In some embodiments, the second gel layer 150 can have a higher concentration of oil than the inner layer.

[0060] In some embodiments, the plant-based scaffold 110, the first gel layer 130, and / or the second gel layer 150 can comprise fibers. In some embodiments, the fibers can comprise animal cells. In some embodiments, the animal cells can comprise myoblasts, mesenchymal stem cells, fibroblasts, keratinocytes, induced pluripotent stem cells, embryonic stem cells, or any combination thereof. In some embodiments, the animal cells can comprise differentiated myotubes and / or adipocytes. In some embodiments, the fibers can comprise cells derived from animal sources, including, but not limited to, domestic cattle, pigs, chickens, quail, and / or rabbits. In some embodiments, the fibers can comprise cells derived from aquatic animals, such as crabs or lobsters. In some embodiments, the fibers can comprise components derived from animal cells.

[0061] In some embodiments, the blended meat product 100 may be heart healthy according to the definition of "heart healthy" provided by the Food and Drug Administration (FDA) pursuant to 21 CFR § 101 (Volume 2). In other words, the blended meat product 100 may be certified with the American Heart Association (AHA) Heart Checkmark. For example, the blended meat product 100 may contain less than 6.5g fat, less than 1g saturated fat (or less than 15% of its calories may come from saturated fat), less than 0.5g trans fat, less than 20mg cholesterol, less than 20mg sodium, and at least 10% of the daily value of at least one of vitamin A, vitamin C, iron, calcium, protein, or dietary fiber per serving (e.g., 50g).

[0062] In some embodiments, the blended meat product 100 may have a hardness value on the food texture characteristic scale of at least about 2N, at least about 2.1N, at least about 2.2N, at least about 2.3N, at least about 2.4N, at least about 2.5N, at least about 2.6N, at least about 2.7N, at least about 2.8N, at least about 2.9N, at least about 3N, at least about 3.1N, at least about 3.2N, at least about 3.3N, at least about 3.4N, at least about 3.5N, at least about 3.6N, at least about 3.7N, at least about 3.8N, or at least about 3.9N. In some embodiments, the blended meat product 100 can have a firmness value of about 4N or less, about 3.9N or less, about 3.8N or less, about 3.7N or less, about 3.6N or less, about 3.5N or less, about 3.4N or less, about 3.3N or less, about 3.2N or less, about 3.1N or less, about 3N or less, about 2.9N or less, about 2.8N or less, about 2.7N or less, about 2.6N or less, about 2.5N or less, about 2.4N or less, about 2.3N or less, about 2.2N or less, or about 2.1 N or less. Combinations of the above firmness values ​​are also possible (e.g., at least about 2N and about 4N or less, or at least about 2.3N and about 3.5N or less), including all values ​​and ranges therebetween. In some embodiments, the blended meat product 100 can have a hardness value of about 2N, about 2.1N, about 2.2N, about 2.3N, about 2.4N, about 2.5N, about 2.6N, about 2.7N, about 2.8N, about 2.9N, about 3N, about 3.1N, about 3.2N, about 3.3N, about 3.4N, about 3.5N, about 3.6N, about 3.7N, about 3.8N, about 3.9N, or about 4N.

[0063] In some embodiments, the blended meat product 100 may have a springiness value on the food texture characteristic scale of at least about 6N, at least about 6.1N, at least about 6.2N, at least about 6.3N, at least about 6.4N, at least about 6.5N, at least about 6.6N, at least about 6.7N, at least about 6.8N, or at least about 6.9N. In some embodiments, the blended meat product 100 may have a springiness value of about 7N or less, about 6.9N or less, about 6.8N or less, about 6.7N or less, about 6.6N or less, about 6.5N or less, about 6.4N or less, about 6.3N or less, about 6.2N or less, or about 6.1N or less. Combinations of the above springiness values ​​(e.g., at least about 6N and about 7N or less, or at least about 6.1N and about 6.9N or less) are also possible, including all values ​​and ranges therebetween. In some embodiments, the blended meat product 100 may have a springiness value of about 6N, about 6.1N, about 6.2N, about 6.3N, about 6.4N, about 6.5N, about 6.6N, about 6.7N, about 6.8N, about 6.9N, or about 7N.

[0064] In some embodiments, the blended meat product 100 may have a cohesiveness value on the food texture attribute scale of at least about 0.4, at least about 0.41, at least about 0.42, at least about 0.43, at least about 0.44, at least about 0.45, at least about 0.46, at least about 0.47, at least about 0.48, at least about 0.49, at least about 0.5, at least about 0.51, at least about 0.52, at least about 0.53, at least about 0.54, at least about 0.55, at least about 0.56, at least about 0.57, at least about 0.58, or at least about 0.59. In some embodiments, the fibrous food products can have a cohesiveness value of about 0.6 or less, about 0.59 or less, about 0.58 or less, about 0.57 or less, about 0.56 or less, about 0.55 or less, about 0.54 or less, about 0.53 or less, about 0.52 or less, about 0.51 or less, about 0.5 or less, about 0.49 or less, about 0.48 or less, about 0.47 or less, about 0.46 or less, about 0.45 or less, about 0.44 or less, about 0.43 or less, about 0.42 or less, or about 0.41 or less. Combinations of the above cohesiveness values ​​(e.g., at least about 0.4 and about 0.6 or less, or at least about 0.45 and about 0.55 or less) are also possible, including all values ​​and ranges therebetween. In some embodiments, the blended meat product 100 may have a cohesiveness value of about 0.4, about 0.41, about 0.42, about 0.43, about 0.44, about 0.45, about 0.46, about 0.47, about 0.48, about 0.49, about 0.5, about 0.51, about 0.52, about 0.53, about 0.54, about 0.55, about 0.56, about 0.57, about 0.58, about 0.59, or about 0.6.

[0065] In some embodiments, the blended meat product 100 may have a stickiness value on the food texture attribute scale of at least about 1, at least about 1.1, at least about 1.2, at least about 1.3, at least about 1.4, at least about 1.5, at least about 1.6, at least about 1.7, at least about 1.8, or at least about 1.9. In some embodiments, the blended meat product 100 may have a stickiness value of about 2 or less, about 1.9 or less, about 1.8 or less, about 1.7 or less, about 1.6 or less, about 1.5 or less, about 1.4 or less, about 1.3 or less, about 1.2 or less, or about 1.1 or less. Combinations of the above stickiness values ​​(e.g., at least about 1 and about 2 or less, or at least about 1.1 and about 1.9 or less) are also possible, including all values ​​and ranges therebetween. In some embodiments, the blended meat product 100 may have a stickiness value of about 1, about 1.1, about 1.2, about 1.3, about 1.4, about 1.5, about 1.6, about 1.7, about 1.8, about 1.9, or about 2.

[0066] In some embodiments, the blended meat product 100 may have a chewiness value on the food texture attribute scale of at least about 0.5, at least about 0.6, at least about 0.7, at least about 0.8, at least about 0.9, at least about 1, at least about 1.1, at least about 1.2, at least about 1.3, or at least about 1.4. In some embodiments, the blended meat product 100 may have a chewiness value of about 1.5 or less, about 1.4 or less, about 1.3 or less, about 1.2 or less, about 1.1 or less, about 1 or less, about 0.9 or less, about 0.8 or less, about 0.7 or less, or about 0.6 or less. Combinations of the above chewiness values ​​(e.g., at least about 0.5 and about 1.5 or less, or at least about 0.6 and about 1.3 or less) are also possible, including all values ​​and ranges therebetween. In some embodiments, the blended meat product 100 may have a chewiness value of about 0.5, about 0.6, about 0.7, about 0.8, about 0.9, about 1, about 1.1, about 1.2, about 1.3, about 1.4, or about 1.5.

[0067] In some embodiments, the blended meat product 100 may have a Warner-Bratzler shear strength of at least about 0.25 kg, at least about 0.5 kg, at least about 1 kg, at least about 1.5 kg, at least about 2 kg, at least about 2.5 kg, at least about 3 kg, at least about 3.5 kg, at least about 4 kg, at least about 4.5 kg, at least about 5 kg, or at least about 5.5 kg. In some embodiments, the blended meat product 100 may have a Warner-Bratzler shear strength of about 6 kg or less, about 5.5 kg or less, about 5 kg or less, about 4.5 kg or less, about 4 kg or less, about 3.5 kg or less, about 3 kg or less, about 2.5 kg or less, about 2 kg or less, about 1.5 kg or less, about 1 kg or less, or about 0.5 kg or less. Combinations of the above Warner-Bratzler shear strengths are also possible (e.g., at least about 0.25 kg and not more than about 6 kg, or at least about 0.5 kg and not more than about 5 kg), including all values ​​and ranges therebetween. In some embodiments, the blended meat product 100 may have a Warner-Bratzler shear strength of about 0.25 kg, about 0.5 kg, about 1 kg, about 1.5 kg, about 2 kg, about 2.5 kg, about 3 kg, about 3.5 kg, about 4 kg, about 4.5 kg, about 5 kg, about 5.5 kg, or about 6 kg.

[0068] 3A-3C are diagrams of a method of making a blended meat product 200, according to one embodiment. FIG. 3A shows a plant-based scaffold 210 placed in a container and submerged in a first carrier material 230a. A second carrier material (not shown) may be added sequentially after the first carrier material 230a. In some embodiments, the plant-based scaffold 210, first carrier material 230a, and second carrier material may be the same as or substantially similar to the plant-based scaffold 110, first carrier material, and second carrier material described above with reference to FIG. 1. Accordingly, certain aspects of the plant-based scaffold 210, first carrier material 230a, and second carrier material will not be described in greater detail herein.

[0069] In some embodiments, the first carrier material 230a and the second carrier material are separate phases. In some embodiments, the first carrier material 230a and the second carrier material may be at least partially mixed together (e.g., in a solution or emulsion). As shown, the plant-based scaffold 210 has a tubular shape. In some embodiments, the plant-based scaffold 210 may have a circular or spherical shape. In some embodiments, the plant-based scaffold 210 may have a substantially linear shape. In some embodiments, the first carrier material 230a may have a deeper penetration depth than the second carrier material. In some embodiments, the second carrier material may have a deeper penetration depth than the first carrier material 230a.

[0070] 3B shows the first carrier material 230a entering the plant-based scaffold 210 and expanding the plant-based scaffold 210. In some embodiments, the gelation of the first carrier material 230a can occur at least partially simultaneously with the infiltration of the first carrier material 230a into the plant-based scaffold 210. During the infiltration of the first carrier material 230a into the plant-based scaffold 210, the heterogeneity and concentration gradients between the first carrier material 230a and the plant-based scaffold 210 begin to dissipate. The first carrier material 230a and the plant-based scaffold 210 become a more homogeneous mass.

[0071] FIG. 3C shows the blended meat product 200 in a fully formed state. As shown, the blended meat product 200 includes an inner layer 230b and an outer layer 250. In some embodiments, the inner layer 230b can be formed as a result of homogenization of the plant-based scaffold 210 and the first carrier material 230a. In some embodiments, the outer layer 250 can be formed as a result of gelation and partial penetration of the second carrier material. In some embodiments, the transition from the outer layer 250 to the inner layer 230b can be gradual, so the boundary between the inner layers 230b can be somewhat blurred. In some embodiments, the outer layer 250 can have properties similar to those of a skin layer, while the inner layer 230b can have properties similar to those of bulk meat. In some embodiments, the inner layer 230b and the outer layer 250 can be the same as or substantially similar to the inner layer and second gel layer 150 described above with reference to FIG. 2. In some embodiments, the blended meat product 200 can be formed in a mold. In some embodiments, the molds can have the shape of a cut of meat, such as a chicken breast, rib, loin, round, flank, brisket, shank, fillet, filet mignon, chuck, sirloin, short loin, front shank, rib, porterhouse, nuggets, tenders, chicken fingers, cutlets, or any other suitable shape factor.

[0072] Figures 4A-4D show the immersion of scaffolds with various levels of carrier liquid diffusion. Figure 4A shows a scaffold immersed in carrier liquid with limited diffusion into the scaffold. Figure 4B shows a scaffold immersed in carrier liquid with complete carrier liquid diffusion. Figure 4C shows the scaffold from Figure 4A after it has been cut open. Figure 4D shows the scaffold from Figure 4B after it has been cut open. As shown, carrier liquid with more complete diffusion produces scaffolds with a darker color and a more complete color scheme.

[0073] Figures 5A-5C show detailed views of a plant-based meat substitute product with a transparent skin covering a scaffold. Figure 5A shows a top perspective view of the product with the scaffold visible through the transparent skin. Figure 5B shows a front view of the product, revealing the sponge-like texture of the scaffold. Figure 5C shows a closer view of the front view of the product, revealing the scaffold texture and skin texture in more detail.

[0074] Figures 6A-6B show scaffolds infiltrated by dipping and by injection. Figure 6A shows a scaffold infiltrated by dipping. As shown, the red area is fully infiltrated with carrier liquid, while the pink and white areas in the center are not. Figure 6B shows the white area of ​​the scaffold infused with carrier liquid within the beige area. As shown, the composition of the center of the scaffold in Figure 6B is somewhat heterogeneous, with localized "hot spots" of carrier liquid.

[0075] Figure 7 shows a cross-section of a plant-based meat substitute product that was histologically stained and imaged at high resolution to reveal cells. Cells are shown as small, dark, oval objects visible in the magnified image and are enclosed in dashed boxes. The magnified image shows porcine fibroblasts in the skin area and porcine myoblasts within the product.

[0076] Various concepts may be embodied as one or more methods, at least one example of which has been provided. Actions performed as part of a method may be ordered in any suitable manner. Thus, while shown as sequential actions in the illustrative embodiments, embodiments may be constructed in which actions are performed in an order different from that illustrated, which may include performing some actions simultaneously. In other words, it should be understood that such features are not necessarily limited to a particular order of execution, but rather to any number of threads, processes, services, servers, etc., that may execute sequentially, asynchronously, concurrently, in parallel, simultaneously, synchronously, etc., in a manner consistent with this disclosure. Thus, some of these features may be inconsistent with one another in that they cannot coexist in a single embodiment. Similarly, some features may be applicable to one aspect of the innovation and not to other aspects.

[0077] Additionally, the present disclosure may include other innovations not currently described. The applicants reserve all rights in such innovations, including the right to embody such innovations and to file additional applications, continuations, continuations-in-part, divisional applications, etc. Thereof. Accordingly, it should be understood that the advantages, embodiments, examples, functional, characteristic, logical, operational, organizational, structural, topological, and / or other aspects of the present disclosure should not be construed as limitations on the present disclosure as defined by the embodiments, or limitations on equivalents of the embodiments. Depending on the particular needs and / or characteristics of individual and / or business users, database organization and / or relational models, data types, data transmission and / or network frameworks, syntax structures, etc., various embodiments of the technology disclosed herein may be implemented in a manner that allows for great flexibility and customization, as described herein.

[0078] All definitions defined and used herein should be understood to supersede dictionary definitions, definitions in documents incorporated by reference, and / or ordinary meanings of the defined terms.

[0079] As used herein, in certain embodiments, the terms "about" or "approximately," when preceding a numerical value, indicate a range of that value ±10%. When a range of values ​​is provided, it is understood that each intervening value between the upper and lower limits of that range, to one-tenth of the unit of the lower limit unless the context clearly dictates otherwise, and any other stated or intervening value within that stated range, is encompassed within the disclosure. The upper and lower limits of these smaller ranges, which may independently be included in the smaller ranges, are also encompassed within the disclosure, subject to any specifically excluded limit in the stated range. When a stated range includes one or both of the limits, ranges excluding either or both of those included limits are also included within the disclosure.

[0080] The term "and / or," as used in the specification and embodiments, should be understood to mean "either or both" of the elements so conjoined, i.e., elements present conjunctively in some cases and disjunctively in other cases. Multiple elements listed with "and / or" should be construed in the same manner, i.e., "one or more" of the elements so conjoined. Other elements, whether related or unrelated to those elements specifically identified, may optionally be present other than the elements specifically identified by the "and / or" clause. Thus, as a non-limiting example, a reference to "A and / or B," when used in conjunction with open-ended language such as "comprising," may refer in one embodiment to A only (optionally including elements other than B); in another embodiment, to B only (optionally including elements other than A); in yet another embodiment, to both A and B (optionally including other elements); and so forth.

[0081] As used in the specification and embodiments, "or" should be understood to have the same meaning as "and / or" as defined above. For example, when separating items in a list, "or" or "and / or" shall be interpreted as being inclusive, i.e., including at least one of, but also including more than one of, several elements or a list of elements, and optionally including additional unlisted items. Only terms clearly indicated to the contrary, such as "only one of" or "exactly one of," or, when used in the embodiments, "consisting of," shall refer to the inclusion of exactly one element of several elements or a list of elements. Generally, as used herein, the term "or" shall be interpreted as indicating exclusive alternatives (i.e., "one or the other but not both") only when preceded by terms of exclusivity, such as "either," "one of," "only one of," or "exactly one of." "Consisting essentially of," when used in the embodiments, shall have its ordinary meaning as used in the field of patent law.

[0082] As used herein and in the embodiments, the phrase "at least one" in reference to a list of one or more elements should be understood to mean at least one element selected from any one or more of the elements in the list of elements, but not necessarily including at least one of each and every element specifically listed in the list of elements, and not excluding any combination of elements in the list of elements. This definition also allows for elements other than those specifically identified in the list of elements to which the phrase "at least one" refers, whether related or unrelated to those specifically identified elements, may optionally be present. Thus, as a non-limiting example, "at least one of A and B" (or equivalently, "at least one of A or B," or equivalently, "at least one of A and / or B") can refer in one embodiment to at least one A, optionally including more than one, with no B present (and optionally including elements other than B); in another embodiment, to at least one B, optionally including more than one, with no A present (and optionally including elements other than A); in yet another embodiment, to at least one A, optionally including more than one, and at least one B, optionally including more than one (and optionally including other elements);

[0083] In the embodiments, as well as in the above specification, all transitional phrases such as "comprise," "include," "carry," "have," "contain," "accompany," "hold," "consisting of," and the like, are to be understood to be open-ended, i.e., to mean including but not limited to. Only the transitional phrases "consisting of" and "consisting essentially of" shall be closed or semi-closed transitional phrases, respectively, as set forth in the United States Patent Office Manual of Patent Examining Procedures, Section 2111.03.

[0084] While specific embodiments of the present disclosure have been outlined above, many alternatives, modifications, and variations will be apparent to those skilled in the art. Accordingly, the embodiments described herein are intended to be illustrative, not limiting. Various changes may be made without departing from the spirit and scope of the present disclosure. Where the methods and steps described above indicate that certain events occur in a particular order, those skilled in the art, having the benefit of this disclosure, will recognize that the order of certain steps may be changed, and that such changes are in accordance with variations of the present invention. In addition, some of the steps may be performed simultaneously, in a parallel process where possible, or sequentially, as described above. While embodiments have been specifically shown and described, it will be understood that various changes in form and detail may be made.

Claims

1. Providing a plant-based scaffold; soaking the plant-based scaffold in a carrier material containing animal cells; gelling the carrier material to the plant-based scaffold; incubating the carrier material and the plant-based scaffold in a controlled environment to produce a meat substitute product; A method comprising:

2. The method of claim 1 , wherein the carrier material comprises a carrier liquid.

3. 3. The method of claim 2, wherein immersing the plant-based scaffold in the carrier material comprises injecting the carrier liquid into the plant-based scaffold.

4. 10. The method of claim 1, wherein the animal cells attach to the plant-based scaffold via an attachment substrate.

5. The method of claim 1 , wherein the animal cells comprise at least one of skeletal muscle cells, fat cells, connective tissue cells, or skin cells.

6. 10. The method of claim 1, wherein the plant-based scaffold has a moisture content of about 40% to about 90% by weight prior to immersing the plant-based scaffold in the carrier material.

7. 10. The method of claim 1, wherein said gelling is by temperature treatment at a temperature of about 60°C to about 120°C.

8. the support material is a first support material; The method comprises:

10. The method of claim 1, further comprising immersing the plant-based scaffold in a second carrier material having a viscosity different from that of the first carrier material and having a penetration depth into the plant-based scaffold different from the penetration depth of the first carrier material.

9. The method of claim 8 , wherein the second carrier material includes a gelling agent and is configured to form a layer of material separate from the first carrier material.

10. 10. The method of claim 1, wherein the controlled environment is at a temperature of less than about 38°C and a pH of about 6.5 to about 8.

11. The method of claim 2 , wherein the carrier liquid undergoes ionic gelation upon addition of the gelling agent.

12. The method of claim 2 , wherein the carrier liquid undergoes gelation upon heating.

13. 10. The method of claim 1, further comprising immersing the plant-based scaffold in a liquid to promote gelation.

14. The method of claim 1 , wherein the carrier material comprises a fat substitute.

15. 10. The method of claim 1, further comprising seeding said plant-based scaffold with animal cells.

16. Plant-based scaffolding and a layer of gel surrounding the plant-based scaffold; the gel layer contains animal cells; at least a portion of the layer of gel permeates the plant-based scaffold; The plant-based scaffold and the gel layer form a composition having organoleptic properties that are the same as or substantially similar to those of meat.

17. 17. The meat substitute product of claim 16, further comprising a fiber comprising at least one of an animal cell or a component derived from an animal cell.

18. the gel layer is a first gel layer; The meat substitute product is 17. The meat substitute product of claim 16, further comprising a second gel layer at least partially surrounding the first gel layer and having a different composition than the first gel layer.

19. 20. The meat substitute product of claim 18, wherein at least one of the plant-based scaffold, the first gel layer, or the second gel layer comprises fiber.

20. 17. The meat substitute product of claim 16, wherein the animal cells comprise at least one of skeletal muscle cells, fat cells, connective tissue cells, or skin cells.

21. 17. The meat substitute product of claim 16, wherein the gel expands the plant-based scaffold.

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