Plant and animal cell mixed meat product and production method thereof

Abstract

Embodiments described herein relate to mixed meat substitutes and methods of producing the same. In some aspects, a method of producing a meat substitute may include providing a plant-based scaffold having a carrier material comprising animal cells, adding a gelling agent to at least one plant-based scaffold, and incubating the carrier material and the plant-based scaffold in a controlled environment to produce the meat substitute. In some embodiments, the animal cells may include at least one of skeletal muscle cells, adipose cells, connective tissue cells, or skin cells.

Description

[0001] Statement Regarding Federally Funded Research

[0002] This invention was made with government support under Grant No. 2112169 awarded by the National Science Foundation. The government has certain rights in the invention.

[0003] Cross - Reference to Related Applications

[0004] This application claims priority to U.S. Provisional Application No. 63 / 407,472, filed on September 16, 2022, titled "Plant and Animal Cell Blended Meat Products and Methods of Producing the Same", the disclosure of which is hereby incorporated by reference in its entirety. Technical Field

[0005] The embodiments described herein relate to plant - based and animal - cell - based meat products and methods of producing the same.

[0006] Background

[0007] Plant - based meats generally have high nutritional value and pose minimal health risks. For example, they can contain any essential amino acids. However, they do not contain the same proteins or fats found in animal tissues and generally lack the sensory characteristics associated with meat from animals. Cell - based meats have recently emerged as a viable alternative to plant - based meats. Cell - based meats contain cultured animal cells, and the animal cells are used to construct muscle and / or fat tissues similar to those from animals. Scaling up cell - based meats is difficult and involves many obstacles. Combining the positive aspects of plant - based meats and cell - based meats can result in products with sensory characteristics similar to those of meats from live animals, which can be produced at a competitive price point.

[0008] Summary

[0009] The embodiments described herein relate to hybrid meat substitutes and methods of producing the same. In some aspects, a method of producing a meat substitute may include providing a plant-based scaffold having a carrier material, the carrier material comprising animal cells or compounds associated with a recombinantly produced animal flavor, 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 a meat substitute. In some embodiments, the animal cells may include at least one of skeletal muscle cells, adipocytes, connective tissue cells, or skin cells. In some embodiments, the carrier may include a carrier fluid. In some embodiments, the carrier material may be a first carrier material, and the method may further include impregnating the plant-based scaffold with a second carrier material having a viscosity different from the first carrier material and a penetration depth in the plant-based scaffold different from the penetration depth of the first carrier material. In some embodiments, the carrier material may be disposed within the scaffold by temperature treatment. In some embodiments, the carrier material may be disposed within the scaffold by ionic gelation. In some embodiments, cells may be cultured in the scaffold. In some embodiments, the penetration depth of the carrier material into the scaffold may be controlled by adjusting the water content of the scaffold. Brief Description of the Drawings

[0011] The patent or application file contains at least one color drawing. Copies of this patent or patent application publication with one or more color drawings will be provided by the Office upon request and payment of the necessary fee.

[0012] Optional features in all figures are shown in dashed lines.

[0013] Figure 1 is a block diagram of a method of producing a hybrid meat product of plant and animal cells according to an embodiment.

[0014] Figure 2 is a block diagram of a hybrid meat product according to an embodiment.

[0015] Figures 3A - 3C is an illustration of a process of producing a hybrid meat product according to an embodiment.

[0016] Figures 4A - 4D shows scaffold soaking with various carrier fluid diffusion levels.

[0017] Figures 5A - 5C shows a detailed view of a plant-based meat substitute having a transparent skin covering the scaffold.

[0018] Figures 6A - 6B shows scaffolds infiltrated by soaking versus injection.

[0019] Figure 7 is an image of a plant-based meat substitute.

[0020] Detailed Description

[0021] The field of tissue engineering encompasses biocompatible materials that serve as scaffolds for the growth of biological cells and tissues. The cell types that form meat include muscle myofibers and adipocytes. Such cells are typically adherent-dependent, such that they rely on attachment substrates for survival and proliferation. To this end, various attachment substrates have been developed over the past few decades. These attachment substrates range from small spherical carriers used in suspension bioreactors to porous 3D matrices used in reaction beds. While there are many applications of tissue engineering, they are mostly limited to medical applications and rarely aim at producing edible products.

[0022] Applications for producing edible products are generally divided into two categories: (1) conventional tissue engineering methods, where cells are cultured within scaffolds, or (2) hybrid products, where cells are propagated using various culture methods and subsequently combined with separate materials that add structure to the final product. Tissue engineering methods typically involve the long-term culture of cells (i.e., for days to weeks, sometimes months) usually within a material scaffold. This allows the cells to mature and develop into dense tissue. Due to diffusion-limited nutrient transport, such methods are typically carried out on a small scale with tissue thicknesses limited to less than 0.5 mm. Efforts to vascularize tissue are underway but have not yet reached the scale or cost for thick tissue production relevant to food production. Feasible tissue engineering applications in the near term include those focused on thin tissues such as skin, cornea, or thin tissues that coat medical devices to improve implant outcomes.

[0023] Methods of making hybrid products have historically involved the expansion of cells and their subsequent addition to a support material. These methods approximate the comparison price of meat from live animals, as the percentage of cells representing the final product depends on the circumstances and may be low compared to most tissue engineering methods. Cells are typically harvested as a dense precipitate and resuspended in a scaffold at a lower density than the original cell precipitate, animal meat tissue, or tissue engineering products that have been continuously cultured. Significant cell proliferation is allowed during culturing. Hybrid products utilize an edible substrate into which cells can be included as an additive to improve nutrition, aroma, and / or flavor. Various methods are used to produce hybrid products to retain cells within their support material. However, when simply added to the support material in an aqueous solution, the distribution of cells within the material is difficult to control, and the cells typically simply flow through the material without being properly retained. Carrier fluids and gelling agents can help retain cells within the support material. Most natural tissues used in meat from live animals have a unique arrangement of multiple cell types that give rise to the characteristic properties of each meat. Hybrid products may also suffer from cell immaturity as they are not given enough time to develop within a structural framework that guides them along the paths seen in natural tissues. For this reason, long muscle fibers are not likely to be present in hybrid products at the prior art level.

[0024] Fibrous plant-based scaffolds can replicate long muscle fiber morphology in terms of length and diameter. Thus, fibrous plant-based scaffolds can contribute to the texture (sensory property) that would be lacking if immature muscle cells were used. For this reason, if the muscle tissue is immature compared to the muscle tissue of an animal, hybrid products based on fibrous scaffolds can have a better texture than even 100% muscle tissue, as has been the case to date. 3D printing is another strategy for imparting a "fibrous" texture, but is subject to significant throughput limitations as the material extrusion rate is inversely proportional to the extrusion diameter, and muscle fiber diameters range between 10 microns and 150 microns, making 3D printing slow and impractical at this scale even with multiple nozzles.

[0025] The methods described herein involve the combination of cultured cells with plant-based meat products to produce hybrid products containing plant components and animal cell components. The methods described herein facilitate the transfer of cells into the plant-based material and the retention of cells within the plant-based material. Additionally, the transfer of specific cell types to specific regions within the plant-based material can promote the production of layered or stratified tissue. The products produced by the methods described herein can range from substantially plant-based products with few animal cells to products that are primarily based on animal cells. In some embodiments, the cells can reproduce within the plant-based material. The embodiments described herein can result in a comparison price with meat from live animals.

[0026] Some embodiments described herein may include the plant proteins and animal cells described in U.S. Provisional Patent Application No. 63 / 346,172, titled "Plant-Based Shredded Meat Products, and Meat Products, and Methods of Producing the Same," filed on May 26, 2022 ("the '172 application"), the disclosure of which is hereby incorporated by reference in its entirety.

[0027] As used in this specification, unless the context clearly dictates otherwise, the singular forms "a," "an," and "the" include plural referents. Thus, for example, the term "a member" is intended to mean a single member or a combination of members, and the term "a material" is intended to mean one or more materials or a combination thereof.

[0028] When used in conjunction with "cylindrical," "linear," and / or other geometric relationships, the term "substantially" 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 although linearity of the portion is desired, some non-linearity may be present in the "substantially linear" portion. Such non-linearity may be caused by manufacturing tolerances or other practical considerations (e.g., pressure or force applied to the support member). Thus, a geometric construct modified by the term "substantially" includes such geometric properties within plus or minus 5% of the geometric construct. For example, a "substantially linear" portion is a portion where the defined axis or centerline is within plus or minus 5% of being linear.

[0029] As used herein, the terms "a set" and "a plurality" may refer to multiple features or a single feature having multiple parts. For example, when referring to a set of fibers, the set of fibers may be considered an electrode having multiple parts, or the set of electrodes may be considered multiple different fibers. Thus, a set of parts or a plurality of parts may include multiple parts that are continuous or discontinuous with each other. A plurality of particles or materials may also be made of multiple articles produced separately and then joined together (e.g., by mixing, an adhesive, or any suitable method). The term "solidification" with respect to a carrier material may refer to the gelling or hardening of the carrier material, characterized by a sharp increase in viscosity or curing. The carrier solution may be solidified within the scaffold by changing the temperature or adding a gelling agent.

[0030] As used herein, "plant" or "plant-based" can include any non-animal-based material used in food production. In other words, "plant" or "plant-based" is not limited to organisms in the plant kingdom. For example, the "plant-based scaffolds" described herein should be understood to include products derived from fungi, such as mycelium, or plant-like protists, such as seaweed or algae.

[0031] The term "progenitor cell" is used herein to refer to a cell that has a more primitive cell phenotype (e.g., is at an earlier stage of a developmental pathway or progression than a fully differentiated cell) and has a higher degree of potential relative to the cells that can be generated from it by differentiation. Typically, progenitor cells also have significant or very high proliferative potential. Depending on the developmental pathway and the environment in which the cells develop and differentiate, progenitor cells can give rise to multiple different differentiated cell types or a single differentiated cell type.

[0032] As used herein, the term "stem cell" refers to an undifferentiated cell that is capable of proliferating and giving rise to more progenitor cells, which have the ability to generate a large number of mother cells, which in turn are capable of generating differentiated or differentiable daughter cells, which are either terminally differentiated or can mature and / or further differentiate. The daughter cells themselves can be induced to proliferate and produce progeny, which then differentiate into one or more mature cell types while also retaining one or more cells with the developmental potential of the parent. The term "stem cell" refers to a subset of progenitor cells that have the ability or potential to differentiate into a more specialized or differentiated phenotype under certain circumstances and, in some cases, retain the ability to proliferate with little differentiation. In one embodiment, the term stem cell generally refers to a naturally occurring mother cell whose progeny (offspring) become specialized in different directions, typically by differentiation, e.g., by acquiring fully individualized 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 can be derived from pluripotent cells that themselves are derived from pluripotent cells, and so on. Although each of these pluripotent cells can be considered a stem cell, the range of cell types that each can produce can vary quite widely. Some differentiated cells also have the ability to give rise to cells with greater developmental potential. This ability can be natural or artificially induced by treatment with various factors. In many biological instances, stem cells are also "pluripotent" because they can produce progeny of more than one different cell type, but this is not required for "stem-ness". Self-renewal is another classic part of the definition of stem cells. In theory, self-renewal can occur by either of two main mechanisms. Stem cells can divide asymmetrically, where one daughter cell remains in the stem cell state and the other daughter cell expresses some different other specific functions and phenotypes. Alternatively, some stem cells in a population can divide symmetrically into two stem cells, thus maintaining some of the stem cells in the population as a whole while the other cells in the population produce only differentiated progeny. Formally, a cell that starts as a stem cell may progress toward a differentiated phenotype but then "reverses" and re-expresses the stem cell phenotype, a term commonly referred to as "dedifferentiation" or "reprogramming" or "reverse differentiation".

[0033] The term "embryonic stem cell" is used to refer to pluripotent stem cells of the inner cell mass of an embryonic blastocyst (see U.S. Pat. Nos. 5,843,780, 6,200,806, the contents of which are incorporated herein by reference). Such cells can similarly be obtained from the inner cell mass of blastocysts generated by somatic cell nuclear transfer (see, e.g., U.S. Pat. Nos. 5,945,577, 5,994,619, 6,235,970, which are incorporated herein by reference). The unique characteristics of embryonic stem cells define the embryonic stem cell phenotype. Thus, if a cell has one or more of the unique characteristics of an embryonic stem cell such that the cell can be distinguished from other cells, then the cell has the embryonic stem cell phenotype. Exemplary unique embryonic stem cell characteristics include, but are not limited to, gene expression profiles, proliferation capacity, differentiation capacity, karyotype, responsiveness to specific culture conditions, etc.

[0034] The term "adult stem cell" or "ASC" is used to refer to any pluripotent stem cell derived from non-embryonic tissues (including fetal, juvenile, and adult tissues). Stem cells have been isolated from a 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.

[0035] Figure 1 is a block diagram of a method 10 for producing a plant and animal cell mixed meat product according to an embodiment. As shown, method 10 includes providing a plant-based scaffold in step 11. Method 10 optionally includes inoculating animal cells into the plant-based scaffold in step 12, and soaking the plant-based scaffold with 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, and incubating the carrier material and the plant-based scaffold in a controlled environment in step 17 to produce a meat substitute.

[0036] Step 11 includes providing a plant-based scaffold. In some embodiments, the plant-based scaffold may comprise plant fibers. In some embodiments, the plant fibers may include phloem fibers, leaf fibers, plant polysaccharides, starch, β-glucan, cellulose, pectin polysaccharides, and / or seed hair fibers. In some embodiments, the plant fibers may include fibers derived from flax, jute, tossa jute, white jute, kenaf, ramie, roselle, sunn, urena, abaca, cantala, henequen, agave, New Zealand flax, sisal, akund floss, bagasse, bamboo, kapok, coir, cotton, silk-cotton tree, kapok, milkweed floss, or any combination thereof. In some embodiments, the plant-based scaffold may comprise plant proteins. In some embodiments, the plant proteins may include proteins derived from rice, peas, soybeans, barley, hulled barley, beans, fava beans, gluten, tempeh, edamame, lentils, chickpeas, nutritional yeast, spelt, teff, seeds, amaranth, quinoa, spirulina, green peas, oats, Ezekiel bread, wild rice, nuts, chia seeds, mycoprotein, mycelium, or any combination thereof. In some embodiments, the plant proteins may comprise one or more amino acids. In some embodiments, the plant proteins may comprise 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 may comprise oils derived from plants. In some embodiments, the oil may be food-safe. In some embodiments, the oil may be organic. In some embodiments, the oil may include coconut oil, canola oil, flaxseed oil, sunflower oil, soybean oil, corn oil, cottonseed oil, olive oil, palm oil, peanut oil, safflower oil, sesame oil, almond oil, beech nut oil, brazil nut oil, cashew oil, hazelnut oil, macadamia nut oil, mongolian nut oil, pecan oil, pine nut oil, pistachio oil, walnut oil, pumpkin seed oil, or any combination thereof.

[0037] In some embodiments, plant-based scaffolds can be grown or produced in a laboratory. In some embodiments, the plant-based scaffolds can be prefabricated (e.g., purchased from a supplier). In some embodiments, the plant-based scaffolds can be provided in their native shape. In some embodiments, the plant-based scaffolds can be formed into a desired shape to facilitate the diffusion of the carrier material into the plant-based scaffolds. In some embodiments, the plant-based scaffolds can be formed by a mold. In some embodiments, the plant-based scaffolds can be 3D printed. In some embodiments, the plant-based scaffolds can have a tubular, substantially linear shape, spherical, or any other suitable form factor or a combination thereof. Other examples of scaffold form factors are described in U.S. Patent Publication 2020 / 0330644, titled “Methods of Forming Three-Dimensional Tissue Scaffolds Using Biological Fiber Inks and Methods of Use Thereof,” filed on October 16, 2018 (“the ‘644 Publication”). The disclosure of this patent publication is hereby incorporated by reference in its entirety.

[0038] Step 12 is optional and includes inoculating the plant-based scaffold with animal cells. In some embodiments, the animal cells can be inoculated into a fibrous, fungus-derived product. In some embodiments, the fibrous, fungus-derived product can comprise mycelia. 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, mixing them with adipocytes, and covering with skin cells as needed. In some embodiments, Step 12 can include delivering muscle and adipocytes deep into the plant-based scaffold. In some embodiments, the animal cells can be inoculated into the plant-based scaffold by injection. In some embodiments, the animal cells can be inoculated into the plant-based scaffold by soaking. In some embodiments, a carrier containing animal cells can be added to the top surface of the scaffold and allowed to soak and / or infiltrate the scaffold. In some embodiments, the animal cells can include skeletal muscle cells, adipocytes, connective tissue cells, skin cells, or any combination thereof. In some embodiments, the animal cells can include mammalian cells, fish cells, avian muscle myoblasts, myoblasts of mammalian origin, myoblasts of avian origin, myoblasts of fish origin, myosatellites, fibroblasts, adipocytes, endothelial cells, epithelial cells, keratinocytes, stem cells, or any combination thereof. In some embodiments, each cell type can be combined with a specific carrier solution having properties customized to deliver the cells to a specific region within the scaffold.

[0039] Step 13 is optional and includes soaking the plant-based scaffold with a liquid. In some embodiments, soaking the plant-based scaffold can facilitate the penetration of one or more carrier materials (e.g., a carrier solution) into the plant-based scaffold. In some embodiments, soaking the plant-based scaffold with a liquid can facilitate the gelation of the carrier material. In some embodiments, the liquid used to soak the plant-based scaffold may contain salts that 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 salts such that the liquid manipulates the osmotic pressure in the plant-based scaffold and draws salts out of the plant-based scaffold. Drawing salts out of the plant-based scaffold can facilitate the 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 contribute to the ionic gelation of a polysaccharide carrier. In some embodiments, the liquid used to soak the plant-based scaffold may contain calcium lactate, calcium chloride, magnesium lactate, potassium-containing compounds (e.g., potassium chloride), and / or magnesium chloride. In some embodiments, the liquid used to soak the plant-based scaffold can contribute to thermogelation. The hydration state of the plant-based scaffold can be a factor in the ability of the plant-based scaffold to absorb carrier materials and gelling agents. In some embodiments, the liquid used to soak the plant-based scaffold can be maintained at a temperature that induces the gelation of the first carrier material. In some embodiments, the liquid used to soak the plant-based scaffold may contain hydrocolloids, 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.

[0040] In some embodiments, step 13 can produce a plant-based scaffold having a moisture content of at least about 35 wt%, at least about 40 wt%, at least about 45 wt%, at least about 50 wt%, at least about 55 wt%, at least about 60 wt%, at least about 65 wt%, at least about 70 wt%, at least about 75 wt%, at least about 80 wt%, at least about 85 wt%, or at least about 90 wt% (i.e., before the plant-based scaffold is impregnated with the carrier material). In some embodiments, step 13 can produce a plant-based scaffold having a moisture content of no more than about 95 wt%, no more than about 90 wt%, no more than about 85 wt%, no more than about 80 wt%, no more than about 75 wt%, no more than about 70 wt%, no more than about 65 wt%, no more than about 60 wt%, no more than about 55 wt%, no more than about 50 wt%, no more than about 45 wt%, or no more than about 40 wt%. Combinations of the above moisture contents are also possible (e.g., at least about 35 wt% and no more than about 95 wt% or at least about 40 wt% and no more than about 90 wt%), including all values and ranges therebetween. In some embodiments, step 13 can produce a plant-based scaffold having a moisture content of about 35 wt%, about 40 wt%, about 45 wt%, about 50 wt%, about 55 wt%, about 60 wt%, about 65 wt%, about 70 wt%, about 75 wt%, about 80 wt%, about 85 wt%, about 90 wt%, or about 95 wt%.

[0041] Step 14 includes adding a first carrier material to the plant-based scaffold. The first carrier material contains animal cells. In some embodiments, the first carrier material may include a carrier liquid. In some embodiments, the carrier liquid may be injected into the plant-based scaffold. In some embodiments, the carrier liquid may be heated. In some embodiments, the carrier liquid undergoes gelation when heated. In some embodiments, the first carrier material may include a carrier gas. In some embodiments, the first carrier material may include a gel. In some embodiments, the animal cells may be mixed with the first carrier material before adding the first carrier material to the plant-based scaffold. In some embodiments, the animal cells may include skeletal muscle cells, adipocytes, connective tissue cells, skin cells, or any combination thereof. In some embodiments, the animal cells may include 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 may 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 may have a concentration of no more than about 100,000,000 cells / ml, no more than about 50,000,000 cells / ml, no more than about 10,000,000 cells / ml, no more than about 5,000,000 cells / ml, 1,000,000 cells / ml, no more than about 500,000 cells / ml, no more than about 100,000 cells / ml, or no more than about 50,000 cells / ml. Combinations of the above cell concentrations are also possible (e.g., at least about 50,000 cells / ml and no more than about 100,000,000 cells / ml or at least about 100,000 cells / ml and no more than about 1,000,000 cells / ml), including all values and ranges therebetween. In some embodiments, the animal cells in the first carrier material may 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.

[0042] In some embodiments, the first carrier material can be water-based, alcohol-based, or oil-based. The carrier material can determine the extent of animal cell infiltration into the scaffold. In some embodiments, the first carrier material can undergo ion gelation. In some embodiments, the first carrier material can comprise a polysaccharide, such as pectin, chitosan, alginate, or any combination thereof. A dilute solution of the carrier liquid that undergoes ion gelation can be used for the infiltration of muscle cells deep into the plant-based scaffold, while a concentrated solution can be used to produce skin. In some embodiments, the first carrier material can comprise agar, pectin, alginate, carrageenan, gellan gum, gelatin, modified starch, methylcellulose, hydroxypropyl methylcellulose, or any combination thereof. In some embodiments, the first carrier material can comprise a polysaccharide dissolved in water. In some embodiments, the amount of the polysaccharide dissolved in water can depend on the desired viscosity of the first carrier material.

[0043] In some embodiments, the first carrier material can undergo thermal gelation. In some embodiments, the first carrier material can comprise 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, gellan gum and / or konjac glucomannan can be added to the scaffold as thermosetting polymers. In some embodiments, the first carrier material can be water-based. 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 comprise water and ethanol. In some embodiments, the first carrier material can comprise water having a monovalent ion, a divalent ion, a salt of a monovalent ion, a salt of a divalent ion, or any combination thereof. In some embodiments, the monovalent ion and / or divalent ion can be a cation, an anion, or a mixture thereof. In some embodiments, the salt can comprise a proton (or hydronium ion) or a hydroxide ion that regulates the pH. In some embodiments, the first carrier material can be formulated to deliver cells to a predetermined region in and around the plant-based scaffold. The carrier material can be formulated to retain cells and prevent the cells from leaking out of the product. In some embodiments, the first carrier material can be formulated such that any fat contained in the first material melts out, causing the product to sizzle during cooking.

[0044] In some embodiments, the first carrier material can be organic (i.e., related to or derived from living matter). In some embodiments, the first carrier material can be certified as organic, as defined by the United States Department of Agriculture (USDA). In some embodiments, the first carrier material can consist of ingredients and / or their certifying agents produced through a process supervised by the USDA National Organic Program (NOP). In some embodiments, the ingredients of the first carrier material can be produced in accordance with the regulations regarding the organic characteristics of the USDA-certified ingredients. In some embodiments, the components of the first carrier material can be produced using "allowed substances" for organic certification, as specified by the USDA in 7 U.S.C.§205(g). In some embodiments, the first carrier material can contain ingredients that are 100% organic by USDA definition (excluding salts and water) (i.e., the ingredients can meet the standards of the USDA's "100% organic" label). In some embodiments, the first carrier material can contain ingredients that are at least 95% organic by USDA definition (excluding salts and water) (i.e., the ingredients can meet the standards of the USDA's "organic" label). In some embodiments, the first carrier material can contain ingredients that are at least 70% organic by USDA definition (excluding salts and water) (i.e., the ingredients can meet the standards of the USDA's "produced with organic _____ " label).

[0045] 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 specified amount of gelling agent) to control the penetration depth of the first carrier material. In some embodiments, animal cell types can be matched with specific carrier fluids (e.g., muscle cells in a diluted carrier fluid that infiltrates a large scaffold volume, followed by skin cells in a thick carrier fluid that coats the surface of the plant-based scaffold). The penetration depth of the first carrier fluid can be a reliable measure of 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 soaking and / or impregnating 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 thermally cured. 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 gels when exposed to a temperature higher than the mixing temperature. For example, a temperature range of about 50°C to about 120°C can be used to gel various starches.

[0046] In some embodiments, the first carrier material and / or the second carrier 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 carrier material and / or the second carrier material can be heated to a temperature not exceeding about 300°C, not exceeding about 290°C, not exceeding about 280°C, not exceeding about 270°C, not exceeding about 260°C, not exceeding about 250°C, not exceeding about 240°C, not exceeding about 230°C, not exceeding about 220°C, not exceeding about 210°C, not exceeding about 200°C, not exceeding about 190°C, not exceeding about 180°C, not exceeding about 170°C, not exceeding about 160°C, not exceeding about 150°C, not exceeding about 140°C, not exceeding about 130°C, not exceeding about 120°C, not exceeding about 110°C, not exceeding about 100°C, not exceeding about 90°C, not exceeding about 80°C, not exceeding about 70°C, not exceeding about 60°C, not exceeding about 50°C, or not exceeding about 40°C. Combinations of the above temperatures are also possible (e.g., at least about 30°C and not exceeding about 300°C or at least about 50°C and not exceeding about 140°C), including all values and ranges therebetween. In some embodiments, the first carrier material and / or the second carrier material can be heated to 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 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.

[0047] In some embodiments, the first carrier material can be injected and then induced to gel by heating. In some embodiments, the scaffold can be compressed and / or massaged before heating to improve the uniformity of the hydrogel distribution produced by heating the carrier solution. 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 uniformly distributed throughout the scaffold. In some embodiments, the first carrier material and / or the second carrier material can be injected into a local area of the scaffold.

[0048] In some embodiments, animal cells can be attached to the scaffold through an attachment matrix. In other words, animal cells can be cultured under conditions that promote cell attachment. The attachment matrix can significantly reduce the time required for animal cells to attach to the scaffold. In some embodiments, the attachment matrix can include spherical carriers used in suspension bioreactors (e.g., spherical carriers with 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 matrix can be composed of a hydrogel. In some embodiments, the attachment matrix can be composed of the same material as the plant-based scaffold (e.g., a combination of polysaccharides and plant proteins).

[0049] Step 15 includes gelling a first carrier material within the plant-based scaffold. In some embodiments, the gelling can be carried out by a gelling agent. In some embodiments, the gelation can be carried out by ionic gelation. In some embodiments, the gelation can be carried out by heat treatment. In some embodiments, the gelling agent can be added by injection into the plant-based scaffold. When injecting the gelling agent, the diffusion distance of the percentage coverage inside the sample starting from the addition point 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 hydrocolloids, methylcellulose, high-viscosity methylcellulose, methylcellulose E / F / K, κ-carrageenan, ι-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 carrier material can undergo gelation through ion exchange. In some embodiments, the first carrier material can undergo gelation through sodium-calcium exchange.

[0050] In some embodiments, gelation can be carried out by temperature treatment (e.g., heat treatment). In some embodiments, the first carrier material within 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 within the plant-based scaffold can be temperature-treated to a temperature not exceeding about 120°C, not exceeding about 115°C, not exceeding about 110°C, not exceeding about 105°C, not exceeding about 100°C, not exceeding about 95°C, not exceeding about 90°C, not exceeding about 85°C, not exceeding about 80°C, not exceeding about 75°C, not exceeding about 70°C, or not exceeding about 65°C. Combinations of the above temperatures are also possible (e.g., at least about 60°C and not exceeding about 120°C or at least about 80°C and not exceeding about 100°C), including all values and ranges therebetween. In some embodiments, the first carrier material within 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, gelation can be carried out by steam treatment.

[0051] In some embodiments, the gelling agent can soak the plant-based scaffold before adding the carrier material with animal cells (i.e., before step 14). In other words, the plant-based scaffold can be pre-soaked with the gelling agent. The pre-soaking can be carried out such that the first carrier material gels when it comes into contact with the gelling agent present in the scaffold. For example, pre-soaking the plant-based scaffold with calcium lactate or calcium chloride will cause the pectin-based carrier to gel when added to the plant-based scaffold. For this 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 before and after adding the first carrier material and animal cells. In some embodiments, a first gelling agent can be added to the plant-based scaffold before adding the first carrier material, and a second gelling agent can be added to the plant-based scaffold after adding the first carrier material.

[0052] In some embodiments, the plant-based scaffold is not pre-soaked in the gelling agent. In such embodiments, the first carrier material and the animal cells are added to the plant-based scaffold, and the plant-based scaffold (with the first carrier material and the animal cells) is impregnated in the gelling agent. In this case, the concentration of the gelling agent is higher (i.e., at least about 5 wt%, at least about 6 wt%, at least about 7 wt%, at least about 8 wt%, at least about 9 wt% or at least about 10 wt% of the weight of the plant-based scaffold) compared to the pre-soaked case, in order to induce immediate or rapid gelation so that the first carrier material and the animal cells are restricted in situ before they can be washed away.

[0053] In some embodiments, the gelling agent can be added stepwise after each combination of the carrier fluid and the animal cells. For example, a diluted carrier fluid containing muscle cells can be added to the plant-based scaffold and then gelled by adding the gelling agent. Then, a thick carrier fluid containing animal cells (skin) can be added as a scaffold coating, and then the complete product is further gelled.

[0054] In some embodiments, the concentration of the gelling agent in the first carrier material can be 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 concentration of the gelling agent in the first carrier material can be no more than about 10 wt%, no more than about 9 wt%, no more than about 8 wt%, no more than about 7 wt%, no more than about 6 wt%, no more than about 5 wt%, no more than about 4 wt%, no more than about 3 wt%, no more than about 2 wt%, no more than about 1 wt%, no more than about 0.9 wt%, no more than about 0.8 wt%, no more than about 0.7 wt%, no more than about 0.6 wt%, no more than about 0.5 wt%, no more than about 0.4 wt%, no more than about 0.3 wt% or no more than about 0.2 wt%. Combinations of the above concentrations of the gelling agent in the first carrier material are also possible (e.g., at least about 0.1 wt% and no more than about 10 wt% or at least about 0.5 wt% and no more than about 8 wt%), including all values and ranges therebetween. In some embodiments, the concentration of the gelling agent in the first carrier material can be 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%.

[0055] In some embodiments, before adding the gelling agent, the first carrier material can have 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, before adding the gelling agent, the first carrier material can have a viscosity of no more than about 10,000 mPa·s, no more than about 9,000 mPa·s, no more than about 8,000 mPa·s, no more than about 7,000 mPa·s, no more than about 6,000 mPa·s, no more than about 5,000 mPa·s, no more than about 4,000 mPa·s, no more than about 3,000 mPa·s, no more than about 2,000 mPa·s, no more than about 1,000 mPa·s, no more than about 900 mPa·s, no more than about 800 mPa·s, no more than about 700 mPa·s, no more than about 600 mPa·s, no more than about 500 mPa·s, no more than about 400 mPa·s, no more than about 300 mPa·s, no more than about 200 mPa·s, no more than about 100 mPa·s, no more than about 90 mPa·s, no more than about 80 mPa·s, no more than about 70 mPa·s, no more than about 60 mPa·s, no more than about 50 mPa·s, no more than about 40 mPa·s, no more than about 30 mPa·s, no more than about 20 mPa·s, no more than about 10 mPa·s, no more than about 9 mPa·s, no more than about 8 mPa·s, no more than about 7 mPa·s, no more than about 6 mPa·s, no more than about 5 mPa·s, no more than about 4 mPa·s, no more than about 3 mPa·s or no more than about 2 mPa·s.Combinations of viscosities as described above are also possible (e.g., at least about 1 mPa·s and not more than about 10,000 mPa·s or at least about 100 mPa·s and not more than about 1,000 mPa·s), including all values and ranges therebetween. In some embodiments, prior to addition of the gelling agent, the first carrier material may have 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 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.

[0056] The viscosity of the first carrier material can be a function of the desired depth of penetration into the plant-based scaffold. For example, the amount of polysaccharide added to water affects the resulting viscosity. After addition of the gelling agent, the first carrier material becomes a carrier gel. The first carrier material can be exposed to the 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, the gelling 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 solidification can be advantageous in determining the extent to which the first carrier material penetrates into the plant-based scaffold prior to full gelling.

[0057] In some embodiments, after adding the gelling agent, the carrier gel may have 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, 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 the gelling agent is added, the carrier gel may have a viscosity of no more than about 100,000 mPa·s, no more than about 90,000 mPa·s, no more than about 80,000 mPa·s, no more than about 70,000 mPa·s, no more than about 60,000 mPa·s, no more than about 50,000 mPa·s, no more than about 40,000 mPa·s, no more than about 30,000 mPa·s, no more than about 20,000 mPa·s, no more than about 10,000 mPa·s, no more than about 9,000 mPa·s, no more than about 8,000 mPa·s, no more than about 7,000 mPa·s, no more than about 6,000 mPa·s, no more than about 5,000 mPa·s, no more than about 4,000 mPa·s, no more than about 3,000 mPa·s, no more than about 2,000 mPa·s, no more than about 1,000 mPa·s, no more than about 900 mPa·s, no more than about 800 mPa·s, no more than about 700 mPa·s, no more than about 600 mPa·s, no more than about 500 mPa·s, no more than about 400 mPa·s, no more than about 300 mPa·s, no more than about 200 mPa·s, no more than about 100 mPa·s, no more than about 90 mPa·s, no more than about 80 mPa·s, no more than about 70 mPa·s, no more than about 60 mPa·s, no more than about 50 mPa·s, no more than about 40 mPa·s, no more than about 30 mPa·s, or no more than about 20 mPa·s. Combinations of the above viscosities are also possible (e.g., at least about 10 mPa·s and no more than about 100,000 mPa·s or at least about 1,000 mPa·s and no more than about 10,000 mPa·s), including all values and ranges therebetween.In some embodiments, after adding a gelling agent, the carrier gel may have 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, 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.

[0058] Step 16 is optional and includes adding a second carrier material to the plant-based scaffold. In some embodiments, the second carrier material may include a carrier liquid. In some embodiments, the second carrier material may include a carrier gas. In some embodiments, the second carrier material may include a gel. The second carrier material may contain animal cells. In some embodiments, the animal cells may be mixed with the second carrier material prior to adding the second carrier material to the plant-based scaffold. In some embodiments, the second carrier material may be significantly different from the first carrier material. For example, the first carrier material may be water-based and the second carrier material may be oil-based. In some embodiments, the second carrier material may be immiscible with the first carrier material. In some embodiments, the second carrier material may be used to create the outer layer or "skin" of the final product. In some embodiments, the second carrier material may contain fats and / or oils. In some embodiments, the second carrier material may have any of the properties described above for the first carrier material (e.g., "certified organic"). In some embodiments, the second carrier material may contain a gelling agent. In some embodiments, the gelling agent contained in the second carrier material may be the same as the gelling agent contained in the first carrier material. In some embodiments, the first carrier material may contain a first gelling agent and the second carrier material may contain a second gelling agent, where the second gelling agent is different from the first gelling agent. In some embodiments, the second carrier material may 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 may be designed to mimic fat. In some embodiments, the second carrier material may be added to the plant-based scaffold by soaking and / or impregnating the plant-based scaffold in the first carrier material. In some embodiments, the first carrier material may be added to the plant-based scaffold by injecting the first carrier material into the plant-based scaffold.

[0059] In some embodiments, the viscosity of the second carrier material can be greater than the viscosity of the first carrier material. In some embodiments, the second carrier material can have a viscosity of 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, 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 carrier material can have a viscosity of no more than about 50,000 mPa·s, no more than about 40,000 mPa·s, no more than about 30,000 mPa·s, no more than about 20,000 mPa·s, no more than about 10,000 mPa·s, no more than about 9,000 mPa·s, no more than about 8,000 mPa·s, no more than about 7,000 mPa·s, no more than about 6,000 mPa·s, no more than about 5,000 mPa·s, no more than about 4,000 mPa·s, no more than about 3,000 mPa·s, no more than about 2,000 mPa·s, no more than about 1,000 mPa·s, no more than about 900 mPa·s, no more than about 800 mPa·s, no more than about 700 mPa·s, no more than about 600 mPa·s, no more than about 500 mPa·s, no more than about 400 mPa·s, no more than about 300 mPa·s, no more than about 200 mPa·s, no more than about 100 mPa·s, no more than about 90 mPa·s, no more than about 80 mPa·s, no more than about 70 mPa·s, no more than about 60 mPa·s, no more than about 50 mPa·s, no more than about 40 mPa·s, no more than about 30 mPa·s, no more than about 20 mPa·s, no more than about 10 mPa·s, no more than about 9 mPa·s, no more than about 8 mPa·s, no more than about 7 mPa·s, or no more than about 6 mPa·s.Combinations of viscosities as described above are also possible (e.g., at least about 5 mPa·s and not more than about 50,000 mPa·s or at least about 100 mPa·s and not more than about 10,000 mPa·s), including all values and ranges therebetween. In some embodiments, after addition of the gelling agent, the first carrier material may have 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 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.

[0060] In some embodiments, the first carrier material and the second carrier material may have different material compositions. In some embodiments, the first carrier material may be a water-based carrier liquid, while the second carrier material may be oil / fat-based. In some embodiments, the first carrier material may be oil / fat-based, while the second carrier material may be water-based. In some embodiments, the first carrier material and the second carrier material may have similar material compositions, but different concentrations. In some embodiments, the first carrier material may comprise water having a first polysaccharide concentration, while the second carrier material may comprise water having a second polysaccharide concentration, the second polysaccharide composition being greater than the first polysaccharide composition. The different concentrations result in different viscosities and scaffold penetration kinetics. In some embodiments, method 10 may include impregnating the plant-based scaffold with 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 foregoing carrier materials is also possible. For example, the plant-based scaffold may be impregnated in one fat / oil-based carrier and two different pectin-based carriers. In some embodiments, the first carrier material may be delivered by a first method, and the second carrier material may be delivered by a second method. In some embodiments, the first carrier material may be delivered by soaking and / or impregnating the plant-based scaffold in the first carrier material, and the second carrier material may be delivered by injecting the second carrier material into the plant-based scaffold. In some embodiments, the first carrier material may be delivered by injecting the first carrier material into the plant-based scaffold, and the second carrier material may be delivered by soaking and / or impregnating the plant-based scaffold in the second carrier material.

[0061] In some embodiments, the second carrier material may be added to the plant-based scaffold at least partially simultaneously with the first carrier material. In some embodiments, the second carrier material may be added to the plant-based scaffold before the first carrier material. In some embodiments, the second carrier material may be added to the plant-based scaffold after the first carrier material. In some embodiments, the penetration depth of the second carrier material in the plant-based scaffold may be less than that of the first carrier material. In some embodiments, the second carrier material may comprise animal cells. In some embodiments, the second carrier material may include skeletal muscle cells, adipocytes, connective tissue cells, or skin cells. In some embodiments, the second carrier material may include 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 may comprise a first type of cell, the second carrier material may comprise a second type of cell, and the second type of cell may be different from the first type of cell. In some embodiments, the first carrier material and the second carrier material may comprise the same type of cell. In some embodiments, the first carrier material may be mixed with muscle myoblasts and delivered deep into the plant-based scaffold, while the second carrier material may be mixed with skin fibroblasts to form skin concentrated near the outer surface of the blended meat product. In some embodiments, the first carrier material may form fat, the second carrier material may form muscle, and the third carrier material may form skin. In some embodiments, the first carrier material that forms fat may undergo temperature-dependent gelation. In some embodiments, the second carrier material that forms muscle may undergo deep delivery by low-concentration ionic gelation. In some embodiments, the third carrier material that forms skin may undergo high-concentration ionic gelation on the surface of the plant- and animal-cell blended meat product. In some embodiments, the first carrier material may comprise a first type of cell, the second carrier material may comprise a second type of cell, and a third type of cell may be injected into the internal region surrounded by the first type of cell and the second type of cell. In some embodiments, the injection of the third type of cell may be performed before soaking the first type of cell and the second type of cell. In some embodiments, the injection of the third type of cell may be performed after soaking the first type of cell and the second type of cell.

[0062] Step 17 is optional and involves incubating one or more carrier materials and a plant-based scaffold in a controlled environment to produce a meat alternative. In some embodiments, an osmotic pressure and concentration gradient can be established within the plant-based scaffold, and one or more carrier materials can be pulled inward towards the center of the plant-based scaffold to reduce or eliminate the concentration gradient during incubation. For example, the plant-based scaffold can have a higher salt concentration than one or more of the carrier materials, such that during incubation, the liquid from one or more of the carrier materials migrates towards the center of the plant-based scaffold to equalize the salt concentration throughout the product. In some embodiments, muscle, fat, and skin cells are separated within the meat alternative such that they reproduce the tissue structure found in a particular meat cut.

[0063] The presence of animal cells in one or more carrier materials and / or plant-based scaffolds can limit the temperature, pH, and osmotic pressure at which incubation occurs. Accordingly, 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 no more than about 40°C, no more than about 39°C, no more than about 38°C, no more than about 37°C, no more than about 36°C, no more than about 35°C, no more than about 34°C, no more than about 33°C, no more than about 32°C, no more than about 31°C, no more than about 30°C, no more than about 29°C, no more than about 28°C, no more than about 27°C, no more than about 26°C, no more than about 25°C, no more than about 24°C, no more than about 23°C, no more than about 22°C, no more than about 21°C, no more than about 20°C, no more than about 15°C, no more than about 10°C, no more than about 5°C, no more than about 0°C, no more than about -5°C, no more than about -10°C, or no more than about -15°C. Combinations of the above incubation temperatures are also possible (e.g., at least about -20°C and no more than about 40°C or at least about 25°C and no more than about 35°C), 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.

[0064] 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 no more than about 8, no more than about 7.9, no more than about 7.8, no more than about 7.7, no more than about 7.6, no more than about 7.5, no more than about 7.4, no more than about 7.3, no more than about 7.2, no more than about 7.1, no more than about 7, no more than about 6.9, no more than about 6.8, no more than about 6.7, no more than about 6.6, no more than about 6.5, no more than about 6.4, no more than about 6.3, no more than about 6.2, or no more than about 6.1. Combinations of the above pH values are also possible (e.g., at least about 6.5 and no more than about 8 or at least about 7 and no 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.

[0065] In some embodiments, the incubation can last 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 last for a period of no more than about 30 days, no more than about 25 days, no more than about 20 days, no more than about 15 days, no more than about 10 days, no more than about 5 days, no more than about 4 days, no more than about 3 days, no more than about 2 days, no more than about 1 day, no more than about 22 hours, no more than about 20 hours, no more than about 18 hours, no more than about 16 hours, no more than about 14 hours, no more than about 12 hours, no more than about 10 hours, no more than about 9 hours, no more than about 8 hours, no more than about 7 hours, no more than about 6 hours, no more than about 5 hours, no more than about 4 hours, no more than about 3 hours, no more than about 2 hours, or no more than about 1 hour.

[0066] In some embodiments, the blended meat product can be formed without an incubation period. In some embodiments, osmotic pressure can be the primary mechanism for attracting the carrier material to its desired location. Scaffold porosity, carrier concentration and viscosity, and the method of osmosis (e.g., passive diffusion, injection) can be used to control the delivery of the carrier into the plant-based scaffold. When cells are incorporated into the plant-based scaffold, the range of osmotic pressure in which they can survive is narrow.

[0067] Combinations of the above incubation times are also possible (e.g., at least about 30 minutes and no more than about 30 days or at least about 2 hours and no more than about 18 hours), including all values and ranges therebetween. In some embodiments, the incubation can last 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.

[0068] In some embodiments, a flavorant can be added to one or more carrier materials and / or the plant-based scaffold prior to the incubation period. In some embodiments, a flavorant can be added to one or more carrier materials and the plant-based scaffold during the incubation period. In some embodiments, a flavorant can be added to one or more carrier materials and the plant-based scaffold after the incubation period. In some embodiments, the flavorant can include a flavor enhancer. In some embodiments, the flavorant can include a fragrance enhancer. In some embodiments, the flavorant can include one or more spices. In some embodiments, the flavorant can include table salt, black pepper, paprika, oregano, anise, celery seed, cinnamon, catnip, cardamom, caraway seed, sassafras, brown mustard, borage, black pepper, mustard seed, dill seed, bergamot, basil, bay leaf, asafoetida, anise, angelica, allspice, cayenne, chervil, chicory, chili, cinnamon bark, coriander leaf, clove, coriander, tansy, curry, dill, fennel, fenugreek, filé, ginger, grains of paradise, holy basil, motherwort, horseradish, hyssop, lavender, lemon balm, lemongrass, lemon verbena, licorice, lovage, mace, oregano, nutmeg, parsley, paprika, parsley, peppermint, rosemary, rue, saffron powder, sage, savory, sesame, sorrel, star anise, spearmint, tarragon, thyme, turmeric, vanilla, wasabi, or any combination thereof. In some embodiments, a colorant can be added before, during, and / or after the incubation period. In some embodiments, animal flavor compounds can be produced from recombinant sources (e.g., recombinant myoglobin) and added to the blended meat product.

[0069] Figure 2 It is a block diagram of a mixed meat product 100 according to an embodiment. As shown, the mixed meat product 100 includes a plant-based scaffold 110 and a first gel layer 130. The mixed 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 sufficiently mixed together such that they jointly form a single-layer material. The combination of the plant-based scaffold 110 and the first gel layer 130 is referred to herein as the "inner layer". In some embodiments, the inner layer can remain separated or partially separated. In some embodiments, the inner layer can form a uniform or substantially uniform single layer.

[0070] In some embodiments, the plant-based scaffold 110 can expand during the formation of the mixed meat product 100. In some embodiments, during the production of the mixed meat product 100, the plant-based scaffold 110 can expand 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. In some embodiments, during the production of the mixed meat product, the plant-based scaffold 110 can become quite soft. In some embodiments, the plant-based scaffold 110 can become harder after gelation. In some embodiments, during the production of the mixed meat product 100, the first gel layer 130 can be included or substantially included in the plant-based scaffold 110. In some embodiments, the penetration depth of the first gel layer 130 within the plant-based scaffold 110, and thus the degree of mixing between the first gel layer 130 and the plant-based scaffold 110, can be controlled by the carrier liquid and gelling agent formulation.

[0071] In some embodiments, the inner layer can have a first composition, the second gel layer 150 can have a second composition, and the second composition is different from the first composition. In some embodiments, the inner layer can have a first texture, the second gel layer 150 can have a second texture, and 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 oil concentration than the inner layer.

[0072] In some embodiments, the plant-based scaffold 110, the first gel layer 130, and / or the second gel layer 150 may comprise fibers. In some embodiments, the fibers may comprise animal cells. In some embodiments, the animal cells may include myoblasts, mesenchymal stem cells, fibroblasts, keratinocytes, induced pluripotent stem cells, embryonic stem cells, or any combination thereof. In some embodiments, the animal cells may include differentiated myotubes and / or adipocytes. In some embodiments, the fibers may comprise cells derived from animal sources (including but not limited to livestock, pigs, chickens, quails, and / or rabbits). In some embodiments, the fibers may comprise cells derived from aquatic animals (such as crabs or lobsters). In some embodiments, the fibers may comprise components derived from animal cells.

[0073] In some embodiments, according to the “heart-healthy” definition provided by the U.S. Food and Drug Administration (FDA) in 21 CFR § 101 (Title 2), the blended meat product 100 may be heart-healthy. In other words, the blended meat product 100 may be certified with the Heart Check mark of the American Heart Association (AHA). For example, each serving (e.g., 50 g) of the blended meat product 100 may contain less than 6.5 g of fat, less than 1 g of saturated fat (or less than 15% of its calories may come from saturated fat), less than 0.5 g of trans fat, less than 20 mg of cholesterol, less than 20 mg of sodium, and at least 10% of the daily value of at least one of vitamin A, vitamin C, iron, calcium, protein, or dietary fiber.

[0074] In some embodiments, the mixed meat product 100 may have a hardness value 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 terms of texture characteristics on a food scale. In some embodiments, the mixed meat product 100 may have a hardness value of no more than about 4N, no more than about 3.9N, no more than about 3.8N, no more than about 3.7N, no more than about 3.6N, no more than about 3.5N, no more than about 3.4N, no more than about 3.3N, no more than about 3.2N, no more than about 3.1N, no more than about 3N, no more than about 2.9N, no more than about 2.8N, no more than about 2.7N, no more than about 2.6N, no more than about 2.5N, no more than about 2.4N, no more than about 2.3N, no more than about 2.2N, or no more than about 2.1N. Combinations of the above hardness values are also possible (e.g., at least about 2N and no more than about 4N or at least about 2.3N and no more than about 3.5N), including all values and ranges therebetween. In some embodiments, the mixed meat product 100 may 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.

[0075] In some embodiments, the mixed meat product 100 may have an elasticity value 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 terms of texture characteristics on a food scale. In some embodiments, the mixed meat product 100 may have an elasticity value of no more than about 7N, no more than about 6.9N, no more than about 6.8N, no more than about 6.7N, no more than about 6.6N, no more than about 6.5N, no more than about 6.4N, no more than about 6.3N, no more than about 6.2N, or no more than about 6.1N. Combinations of the above elasticity values are also possible (e.g., at least about 6N and no more than about 7N or at least about 6.1N and no more than about 6.9N), including all values and ranges therebetween. In some embodiments, the mixed meat product 100 may have an elasticity 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.

[0076] In some embodiments, the mixed meat product 100 may have a gumminess value 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 terms of texture characteristics on a food scale. In some embodiments, the fibrous food may have a gumminess value of no more than about 0.6, no more than about 0.59, no more than about 0.58, no more than about 0.57, no more than about 0.56, no more than about 0.55, no more than about 0.54, no more than about 0.53, no more than about 0.52, no more than about 0.51, no more than about 0.5, no more than about 0.49, no more than about 0.48, no more than about 0.47, no more than about 0.46, no more than about 0.45, no more than about 0.44, no more than about 0.43, no more than about 0.42, or no more than about 0.41. Combinations of the above gumminess values are also possible (e.g., at least about 0.4 and no more than about 0.6 or at least about 0.45 and no more than about 0.55), including all values and ranges therebetween. In some embodiments, the mixed meat product 100 may have a gumminess 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.

[0077] In some embodiments, the mixed meat product 100 may have a gumminess value 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 terms of texture characteristics on a food scale. In some embodiments, the mixed meat product 100 may have a gumminess value of no more than about 2, no more than about 1.9, no more than about 1.8, no more than about 1.7, no more than about 1.6, no more than about 1.5, no more than about 1.4, no more than about 1.3, no more than about 1.2, or no more than about 1.1. Combinations of the above gumminess values are also possible (e.g., at least about 1 and no more than about 2 or at least about 1.1 and no more than about 1.9), including all values and ranges therebetween. In some embodiments, the mixed meat product 100 may have a gumminess 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.

[0078] In some embodiments, the comminuted meat product 100 may have a chewiness value 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 terms of the texture characteristics on a food scale. In some embodiments, the comminuted meat product 100 may have a chewiness value of no more than about 1.5, no more than about 1.4, no more than about 1.3, no more than about 1.2, no more than about 1.1, no more than about 1, no more than about 0.9, no more than about 0.8, no more than about 0.7, or no more than about 0.6. Combinations of the above chewiness values are also possible (e.g., at least about 0.5 and no more than about 1.5 or at least about 0.6 and no more than about 1.3), including all values and the ranges therebetween. In some embodiments, the comminuted 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.

[0079] In some embodiments, the comminuted meat product 100 may have a Warner - Bratzler shear force 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 comminuted meat product 100 may have a Warner - Bratzler shear force of no more than about 6 kg, no more than about 5.5 kg, no more than about 5 kg, no more than about 4.5 kg, no more than about 4 kg, no more than about 3.5 kg, no more than about 3 kg, no more than about 2.5 kg, no more than about 2 kg, no more than about 1.5 kg, no more than about 1 kg, or no more than about 0.5 kg. Combinations of the above Warner - Bratzler shear forces are also possible (e.g., at least about 0.25 kg and no more than about 6 kg or at least about 0.5 kg and no more than about 5 kg), including all values and the ranges therebetween. In some embodiments, the comminuted meat product 100 may have a Warner - Bratzler shear force of about 0.25 kg, 0.5 kg, 1 kg, 1.5 kg, 2 kg, 2.5 kg, 3 kg, 3.5 kg, 4 kg, 4.5 kg, 5 kg, 5.5 kg, or 6 kg.

[0080] Figures 3A - 3C is a diagram of a method for producing a comminuted meat product 200 according to an embodiment. Figure 3AA plant-based scaffold 210 disposed in a container and immersed in a first carrier material 230a is shown. A second carrier material (not shown) may be sequentially added after the first carrier material 230a. In some embodiments, the plant-based scaffold 210, the first carrier material 230a, and the second carrier material may be the same as or substantially similar to the plant-based scaffold 110, the first carrier material, and the second carrier material, as described above with reference to Figure 1 those described. Accordingly, certain aspects of the plant-based scaffold 210, the first carrier material 230a, and the second carrier material are not described in more detail herein.

[0081] 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 greater penetration depth than the second carrier material. In some embodiments, the second carrier material may have a greater penetration depth than the first carrier material 230a.

[0082] Figure 3B The first carrier material 230a entering the plant-based scaffold 210 and causing the plant-based scaffold 210 to expand is shown. In some embodiments, the gelation of the first carrier material 230a may occur at least partially simultaneously with the penetration of the first carrier material 230a into the plant-based scaffold 210. During the penetration of the first carrier material 230a into the plant-based scaffold 210, the heterogeneity and concentration gradient 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 more homogeneous objects.

[0083] Figure 3CShows the commingled meat product 200 in a fully formed state. As shown, the commingled meat product 200 includes an inner layer 230b and an outer layer 250. In some embodiments, the inner layer 230b may be formed due to the homogenization of the plant-based scaffold 210 and the first carrier material 230a. In some embodiments, the outer layer 250 may be formed due to the gelling and partial penetration of the second carrier material. In some embodiments, the boundary between the inner layers 230b may be somewhat blurred because the transition from the outer layer 250 to the inner layer 230b may be gradual. In some embodiments, the outer layer 250 may have properties similar to those of a skin layer, while the inner layer 230b may have properties similar to those of bulk meat. In some embodiments, the inner layer 230b and the outer layer 250 may be the same as or substantially similar to the inner layer and the second gel layer 150, respectively, as described above with reference to Figure 2 Those described. In some embodiments, the commingled meat product 200 may be formed in a mold. In some embodiments, the mold may have the shape of a cut of meat. In some embodiments, the mold may have the shape of a chicken breast, rib, loin, round, flank, brisket, shank, tenderloin, filet mignon, chuck, beef tenderloin, short loin, fore shank, short plate, top sirloin steak, nugget, tender, chicken finger, cutlet, or any other suitable shape factor.

[0084] Figures 4A - 4D Shows scaffold soaking with various levels of carrier fluid diffusion. Figure 4A Shows a scaffold soaked with a carrier fluid that has limited diffusion into the scaffold. Figure 4B Shows a scaffold soaked with a carrier fluid that has complete carrier fluid diffusion. Figure 4C Shows the scaffold after being cut from Figure 4A . Figure 4D Shows the scaffold after being cut from Figure 4B . As shown, a carrier fluid with more complete diffusion results in a scaffold with a darker color and a more thorough coloring scheme.

[0085] Figures 5A - 5C Shows a detailed view of a plant-based meat substitute with a transparent skin covering the scaffold. Figure 5A Shows a top perspective view of the product, where the scaffold is visible through the transparent skin. Figure 5B Shows a front view of the product, where the spongy texture of the scaffold is visible. Figure 5C Shows a closer view of the front view of the product, where more details of the scaffold texture and the skin texture are visible.

[0086] Figures 6A - 6B Shows a scaffold infiltrated by contrast injection through soaking. Figure 6A Shows a scaffold infiltrated by soaking. As shown, the redder parts have been completely infiltrated by the loading solution, while the middle pink and white parts have not been completely infiltrated by the loading solution. Figure 6B Shows the white part of a scaffold in which the loading solution has been injected into the beige part. As shown, Figure 6B the composition in the center of the scaffold is somewhat non-uniform, with local "hot spots" of the loading solution.

[0087] Figure 7 Shows a cross-section of a plant-based meat substitute that has been histologically stained and imaged at high resolution to show cells. In the magnified image, the cells appear as small, dark oval objects, which are surrounded by a dashed box. The magnified view shows porcine fibroblasts in the skin area and porcine myoblasts within the product.

[0088] Various concepts can be embodied as one or more methods, for which at least one example has been provided. The operations performed as part of the method can be sequenced in any suitable way. Accordingly, embodiments can be constructed in which the operations are performed in an order different from that shown, and such embodiments can include performing some operations simultaneously (even if shown as sequential operations in the illustrative embodiments). In other words, it should be understood that such features need not be limited to a particular order of execution, but can be executed serially, asynchronously, concurrently, in parallel, simultaneously, synchronously, and / or similarly, etc., by any number of threads, processes, services, servers, etc. in a manner consistent with the present disclosure. Accordingly, some of these features may be mutually contradictory, as they cannot appear in a single embodiment simultaneously. Similarly, some features apply to one aspect of the innovation and not to other aspects.

[0089] In addition, the present disclosure may include other innovations not currently described. The applicant reserves all rights in such innovations, including the right to implement such innovations, file additional applications, their continuations, partial continuations, divisional applications, etc. Accordingly, it should be understood that the advantageous aspects, embodiments, examples, functions, features, logics, operations, organizations, structures, topologies, and / or other aspects of the present disclosure should not be considered as limitations on the present disclosure defined by the embodiments or on equivalents of the embodiments. Depending on the specific expectations and / or features of individual and / or enterprise users, database configurations and / or relational models, data types, data transmissions, and / or network frameworks, syntactic structures, etc., various embodiments of the techniques disclosed herein can be implemented in a manner that achieves a great deal of flexibility and customization as described herein.

[0090] All definitions defined and used herein should be understood to be based on dictionary definitions, definitions in incorporated-by-reference documents, and / or the ordinary meaning of the defined terms.

[0091] As used herein, in certain embodiments, the term "about" or "substantially" before a numerical value means that value plus or minus a range of 10%. Where a numerical range is provided, it is to be understood that each intervening numerical value between the upper and lower limits of that range (to one-tenth of the lower limit unit, unless otherwise clearly specified herein) as well as any other stated or intervening numerical value within that range is included in the invention. The upper and lower limits of these smaller ranges may independently be included in the smaller ranges and are also included in the present disclosure, subject to any specifically excluded limits within the stated range. When the range includes one or both of the upper and lower limits, ranges excluding one or both of those included limits are also included in the invention.

[0092] The phrase "and / or" as used in the specification and embodiments herein should be understood to mean "either or both" of the elements so conjoined, i.e., elements that exist 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 may optionally exist, whether or not related to those specifically identified, in addition to the elements specifically recited in the "and / or" clause. Thus, as a non-limiting example, in one embodiment, when used in conjunction with open-ended language such as "comprising," a reference to "A and / or B" can refer only to A (optionally including elements other than B); in another embodiment, only to B (optionally including elements other than A); in yet another embodiment, it should refer to both A and B (optionally including other elements); and so forth.

[0093] 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" should be interpreted as inclusive, i.e., including at least one of a plurality or series of elements and optionally additional unlisted items, but also including more than one. Only terms that expressly state the contrary, such as "only one of... " or "exactly one of...," or when used in an embodiment, "consisting of...," will refer to including exactly one element of a plurality or series of elements. In general, the term "or" as used herein should be interpreted as an exclusive alternative only when preceded by exclusive terms such as "either," "one of...," "only one of...," or "exactly one of...," i.e., "one or the other, but not both." When used in an embodiment, "consisting essentially of... " should have its ordinary meaning as used in the field of patent law.

[0094] As used in the specification and the embodiments, the phrase "at least one" with respect 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 specific element listed in the list of elements, and not excluding any combination of elements in the list of elements. This definition also allows that, in addition to the elements specifically identified in the list of elements referred to by the phrase "at least one", elements may optionally exist, whether or not related to those specifically identified elements. 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") may, in one embodiment, mean at least one A, optionally including more than one A, with no B present (and optionally including elements other than B); in another embodiment, it may mean at least one B, optionally including more than one B, with no A present (and optionally including elements other than A); in yet another embodiment, it may mean at least one A, optionally including more than one A, and at least one B, optionally including more than one B (and optionally including other elements); and so on.

[0095] In the embodiments and in the foregoing specification, all transitional phrases such as "comprising", "including", "carrying", "having", "containing", "involving", "holding", "composed of", and the like, are to be understood to be open-ended, i.e., to mean including but not limited to. As set forth in section 2111.03 of the United States Patent and Trademark Office's Manual of Patent Examining Procedure, only the transitional phrases "consisting of" and "consisting essentially of" are closed or semi-closed transitional phrases, respectively.

[0096] Although 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 set forth herein are intended to be illustrative and 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 of ordinary skill in the art who benefit from the present disclosure will recognize that the order of some steps may be modified, and such modifications are in accordance with the variations of the invention. Additionally, where possible, certain steps may be performed simultaneously in a parallel process as well as in the order described above. Specific embodiments have been shown and described in detail, but it should be understood that various changes in form and detail may be made.

Claims

1. A method, the method comprising: Providing a plant-based scaffold; Impregnating the plant-based scaffold with a carrier material, the carrier material comprising animal cells; Gelling the carrier material onto the plant-based scaffold; and Incubating the carrier material and the plant-based scaffold in a controlled environment to produce a meat substitute.

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

3. The method according to claim 2, wherein impregnating the plant-based scaffold with the carrier material comprises injecting the carrier liquid into the plant-based scaffold.

4. The method according to claim 1, wherein the animal cells are attached to the plant-based scaffold via an attachment matrix.

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

6. The method according to claim 1, wherein prior to impregnating the plant-based scaffold with the carrier material, the plant-based scaffold has a moisture content of about 40 wt% to about 90 wt%.

7. The method according to claim 1, wherein the gelling is carried out by a temperature treatment at a temperature between about 60 °C and about 120 °C.

8. The method according to claim 1, wherein the carrier material is a first carrier material, and the method further comprises: Impregnating the plant-based scaffold with a second carrier material having a viscosity different from that of the first carrier material, and the second carrier material having a penetration depth different from that of the first carrier material in the plant-based scaffold.

9. The method according to claim 8, wherein the second carrier material comprises a gelling agent and is configured to form a material layer separated from the first carrier material.

10. The method according to claim 1, wherein the controlled environment is at a temperature below about 38 °C and a pH value between about 6.5 and about 8.

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

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

13. The method according to claim 1, the method further comprises: Soaking the plant-based scaffold with a liquid to facilitate gelling.

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

15. The method according to claim 1, the method further comprises: Inoculating animal cells into the plant-based scaffold.

16. A meat substitute, comprising: A plant-based scaffold; A gel layer surrounding the plant-based scaffold, the gel layer comprising animal cells, at least a portion of the gel layer penetrating the plant-based scaffold, and the plant-based scaffold and the gel layer forming a composition having sensory characteristics the same as or substantially similar to those of meat.

17. The meat substitute according to claim 16, further comprising fibers, the fibers comprising at least one of animal cells or components derived from animal cells.

18. The meat substitute according to claim 16, wherein the gel layer is a first gel layer, and the meat substitute further comprises: A second gel layer that at least partially surrounds the first gel layer, the second gel layer having a composition different from that of the first gel layer.

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

20. The meat substitute according to claim 16, wherein the animal cells comprise at least one of skeletal muscle cells, adipocytes, connective tissue cells, or skin cells.

21. The meat substitute according to claim 16, wherein the gel expands the plant-based scaffold.

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