93 results about "Cultured meat" patented technology

The present invention provides a serum-free culture medium for in vitro suspension culture of porcine muscle stem cells and its application, which can help porcine muscle stem cells expand and be suspended under serum-free and carrier-free conditions. Compared with traditional porcine muscle stem cell culture protocols and culture media, this serum-free suspension culture medium enables porcine muscle stem cells to be cultured continuously at high densities in both batch and fed-batch culture processes under serum-free and carrier-free suspension culture conditions, while maintaining high cell viability and achieving a peak cell density of 1×10 7 The beneficial effect of the present invention is to effectively solve the problem of serum-free and carrier-free efficient suspension expansion of cell cultured meat seed cells, and provide a technical and raw material basis for the industrial production of cell cultured meat.

The invention discloses a porcine muscle stem cell line capable of being cultured in vitro for a long time and application of the porcine muscle stem cell line, and belongs to the technical field of stem cells and cell culture meat. The porcine muscle stem cell line is prepared by the following steps: performing CDKN2A gene knockout on porcine muscle stem cells by using a CRISPR / Cas9 gene editing technology, performing monoclonal screening and genotype identification to obtain a cell strain with successful CDKN2A knockout, and then performing carrier-free and serum-free suspension culture domestication on the cell strain. And a carrier-free and serum-free porcine muscle stem cell line capable of being subjected to in-vitro long-term suspension culture is obtained. Verification shows that the CDKN2A knockout porcine muscle stem cell line subjected to carrier-free and serum-free suspension culture domestication not only can be subjected to carrier-free and serum-free suspension culture in a shake flask for a long time, but also can maintain a good differentiation characteristic. The porcine muscle stem cell line which can be cultured on a large scale, is derived from edible animals and can be cultured in vitro for a long time is created, and excellent cell resources are provided for commercial production of cultured meat.

The invention discloses a porcine fibroblast line capable of being passaged for a long time and application of the porcine fibroblast line. According to the invention, pig fibroblast tissues are taken as a source, pig fibroblasts at different parts are obtained through a tissue block separation method, in-vitro long-term passage is carried out, the cells are triggered to break through the Harike limit, monoclonal screening is further carried out through a limited dilution method, and a pig fibroblast line with a single source is obtained. After screening, porcine fibroblasts with two parts can be subjected to long-term passage in vitro, namely ear edge fibroblasts and rear leg fibroblasts, have collagen secretion capability and are preserved in China Center for Type Culture Collection with preservation numbers of CCTCC NO: C202511 and CCTCC NO: C202512, the cell line breaks through the Harike limit, is already transferred to 42 generations at present, and has the advantages of being capable of secreting collagen, capable of secreting collagen, capable of secreting collagen and capable of secreting collagen. The cells are in a healthy and full long fusiform state, the proliferation level is not influenced after long-term passage, and the method can be applied to preparation of cell culture meat.

The invention provides a serum-free culture medium for in-vitro suspension culture of porcine muscle stem cells and application of the serum-free culture medium. The serum-free culture medium can help the porcine muscle stem cells to be subjected to suspension enlarged culture under serum-free and carrier-free conditions. Compared with a traditional porcine muscle stem cell culture scheme and culture medium, the serum-free suspension culture medium has the advantages that high-density continuous culture of porcine muscle stem cells can be realized in a batch culture process and a fed-batch culture process under the serum-free carrier-free suspension culture condition, high cell viability is maintained, and the survival rate of the porcine muscle stem cells is increased. The peak density of the cells reaches 1 * 10 < 7 > cells / mL or above, and the cell viability is maintained to be 90% or above. The method has the beneficial effects that the problem of serum-free carrier-free efficient suspension amplification of the seed cells of the cell culture meat is effectively solved, and a technical and raw material basis is provided for industrial production of the cell culture meat.

The invention provides a preparation method of a cell culture three-dimensional scaffold and cell culture meat, which comprises the following steps: firstly, preparing a gelatin-carrageenan mixed solution and a gel precursor solution of transglutaminase, dividing into two parts, incubating at constant temperature to obtain two parts of composite hydrogel, randomly freeze-drying one part to obtain aerogel, and freeze-drying the aerogel to obtain the cell culture three-dimensional scaffold. Stacking on the other part and incubating at constant temperature again to obtain the prefabricated double-layer gel. The prefabricated double-layer gel is directionally frozen, ice crystals grow upwards from the bottom, freezing is stopped after the temperature is reduced to the set temperature, and the cell culture three-dimensional scaffold is obtained through drying. The stent adopts an ordered-disordered partition design, parallel micro-channels are formed in the lower layer by using a directional ice template method, and muscle fiber arrangement is simulated; a porous spongy structure is constructed on the upper layer by using a random ice template method, and is adapted to adipocyte proliferation. Two layers of interfaces are interpenetrated and communicated, natural mechanical gradients are combined, natural tissue characteristics are re-engraved, the traditional limitation is broken through, the cell co-culture efficiency and the cultured meat quality are improved, and industrialization is promoted.

A cell-cultured meat production device based on microfluidic 3D printing technology includes a printing nozzle, a printing moving system, a loading platform, a sample injection system and a base. The printing moving system is disposed on the base and is composed of a plurality of movable optical axes. The printing nozzle is disposed on a movable optical axis, the sample injection system is connected to the printing nozzle, and the printing nozzle is a micro-fluidic chip. The present invention is based on microfluidic 3D printing, where biological ink is extruded by means of a microfluidic chip to form printed fibers that, under the drive of the printing moving system, are stacked for formation to achieve single-step construction of a piece of three-dimensional tissue having tissue anisotropy, which is used for cell-cultured meat production.

The present invention belongs to the field of cell culture media. Provided are a method for preparing a stem cell culture medium by using a protein hydrolysate as a core raw material and a method of using the stem cell culture medium. The stem cell culture medium contains 20-80% of the protein hydrolysate on the basis of the total weight of the components in the stem cell culture medium. The protein hydrolysate comprises one or more of a high-free amino acid yeast hydrolysate, a high-nucleotide yeast hydrolysate, a high-glutathione yeast hydrolysate, a high-polypeptide yeast hydrolysate, a rice protein hydrolysate, a high-glutamine wheat protein hydrolysate, a pea protein hydrolysate and a soybean protein hydrolysate. The preparation of a natural animal stem cell culture medium by using a whole nutrient protein hydrolysate as a core raw material can meet the nutritional requirements for animal stem cell proliferation under the conditions of no additional addition or addition of a small amount of amino acids, vitamins, growth factors and serum, which effectively reduces the cost of the stem cell culture medium, thereby providing the stem cell culture medium with wide application prospects in the field of foods such as cultured meat.

The present invention discloses a low - serum medium and its application in the preparation of cultured meat from Larimichthys crocea cells, which is also applicable to other marine or freshwater fish, belonging to the field of bioengineering technology. The low - serum medium comprises a basal medium, fetal bovine serum accounting for 30 - 80 μL / mL, and exogenous additives. The exogenous additives include human serum albumin, ascorbic acid, basic fibroblast growth factor polypeptide, human holo - transferrin, lysophosphatidic acid, and human recombinant IGF - 1. Based on the DMEM / F12 medium, the low - serum medium provided by the present invention adds six substances that promote cell growth, reduces the dependence of satellite cells on fetal bovine serum, effectively reduces the usage amount of fetal bovine serum, and saves the culture cost. Compared with the general - type medium (DMEM high - glucose, adding 10% FBS), the low - serum medium provided by the present invention has an equivalent effect on the maintenance and proliferation ability of cell stem cells.

The objective of the present invention is to provide a method for manufacturing powdered cultured meat that can increase cell differentiation efficiency and protein content with economic feasibility, and to provide powdered cultured meat with high physicochemical stability and transportability, as well as a food composition containing the same. The powdered cultured meat has a flavor similar to real meat and, as a high-protein food, has excellent utility in various food compositions or processed foods.

The present invention discloses a porcine fibroblast cell line capable of long-term passage and its application. The present invention uses porcine fibroblast tissue as a source, obtains porcine fibroblasts from different parts through tissue block separation, performs long-term in vitro passage, and induces the cells to break through the Hayflick limit. Further, monoclonal screening is performed through limiting dilution to obtain a single-source porcine fibroblast cell line. After screening, porcine fibroblasts from two parts, ear margin fibroblasts and hind leg fibroblasts, can be long-term passaged in vitro and have the ability to secrete collagen. These cells have been deposited with the China Center for Type Culture Collection under the accession numbers CCTCC NO: C202511 and CCTCC NO: C202512. This cell line has broken through the Hayflick limit and has been passaged to 42 generations. The cells exhibit a healthy, plump, long spindle shape. Long-term passage does not affect proliferation levels, and the cell line can be used in the preparation of cell-cultured meat.

The present disclosure provides extruded or spun semi-permeable porous hollow fibers comprising covalent ester, thioester, and / or amide cross-linked polypeptides and methods of producing the same. The hollow fibers may be produced from proteins, protein extracts and / or protein isolates derived from plants, animals, bacteria, algae, archaea and / or fungi and, in certain embodiments, are intended to be suitable for human and / or animal ingestion. In some embodiments, the hollow fibers may be designed for producing cartridges compatible with existing and / or novel bioreactor platforms for containing cell cultures in cultured meat production.

The present disclosure relates to cultured tissue, methods for production of the cultured tissue, e.g., cultured meat, that is not genetically modified. Further, the disclosure provides method of temporary immortalization of cells for propagation in vitro.

The invention discloses a composite modifier and a method for improving the texture of meat paste of cultured meat, belongs to the technical field of novel food processing, and aims to solve the problems of loose structure, poor chewing feeling, insufficient water-retaining property and the like of the cultured meat. The method comprises the following steps: chopping and mixing a composite modifier containing soybean protein isolate and distarch phosphate in a mass ratio of (1-9): (9-1) with a culture meat paste base material at 4 DEG C or below according to a proportion of 5-15% of the total weight of the culture meat paste, then carrying out centrifugal degassing on the mixed meat paste, carrying out water bath heating gelation at 80-90 DEG C, and carrying out cooling storage at 2-6 DEG C, therefore, a stable gel network with enhanced hardness, elasticity and water binding capacity is constructed, and the gel network is mainly used for improving the texture quality and sensory acceptance of cultured meat products.

This invention discloses a multi-enzyme preparation for industrial cell dissociation, its usage method, and applications, belonging to the field of seed cell extraction technology for industrial meat culture. The invention employs a multi-enzyme preparation composed of component 1 and component 2; component 1 includes neutral protease, hyaluronidase, and elastase; component 2 includes matrix metalloproteinase, papain, and plasmin. The invention also discloses the preparation method and applications of the multi-enzyme preparation. The rational enzyme ratio and optimized treatment conditions of this invention help protect cell membrane integrity, maintain high cell activity, and significantly improve dissociation efficiency and cell recovery. The compound enzyme preparation is highly adaptable, applicable to various tissue types and dissociation requirements, and facilitates standardized and large-scale production. Combined with automated dissociation equipment, the compound enzyme preparation enables efficient and stable industrial operation, providing important technical support and broad application prospects for the cultured meat industry, tissue engineering, and cell therapy.

A new cell culture system capable of improving the efficiency in cell culture is provided. A cell culture system comprises a plurality of cell culture devices which are connected such that a culture medium moves through each of the cell culture devices, and a liquid level of the culture medium stored in each of the cell culture devices is controlled such that cells in at least one of the cell culture devices are in a first state in which the cells are immersed in the culture medium, while cells in the remaining cell culture devices are in a second state in which the cells are not immersed in the culture medium.

The present invention relates to: a cultured meat medium additive and a cultured meat medium composition that can induce muscle growth by differentiating muscle stem cells into muscle cells and inhibiting the expression of myostatin, which inhibits muscle growth; and a method for preparing cultured meat by using same. Specifically, the cultured meat medium additive and the cultured meat medium composition comprise laxogenin and 5-alpha hydroxy laxogenin.

The present invention relates to a single-phase, spherical edible microcarrier for the three-dimensional culture of stem cells intended for the preparation of cultured meat. The microcarrier comprises a homogeneous mixture of edible ionic polysaccharide (0.375-1.5%), edible protein (1-2.5%), edible polymer (0.025-0.1%) and water (95.9-98.6%). The invention also relates to a method for preparing these microcarriers by mixing three solutions: ionic polysaccharide in saline solution, edible protein in saline solution and edible polymer in water. The mixture is subjected to a process involving microspraying or electrospraying with calcium chloride in order to form spherical microcarriers of 50-300 micrometres. The design of the single-phase, spherical edible microcarrier simplifies the production of cultured meat by dispensing with the need for core-shell structures, thereby improving cell anchorage efficiency and facilitating cell recovery.

The invention provides a method for culturing larimichthys crocea cells as animal and plant composite food by a one-step method with rice grains as a support, and belongs to the technical field of food biology. The invention relates to application of rice as a scaffold in preparation of cell culture meat from pseudosciaena crocea cells. Experimental results show that myoblasts and / or precursor adipocytes of the pseudosciaena crocea can normally proliferate on the rice grain scaffold, the rice nutrition is guaranteed, the flavor similar to that of real fish meat is achieved, and the prepared cell culture meat has the potential of being developed into sustainable future food.

This invention relates to the fields of stem cell culture and serum-free culture media, specifically to a serum-free culture medium for the proliferation of bovine muscle stem cells and its applications. The serum-free culture medium is based on a serum-free basal culture medium, to which food-derived small molecule compounds are added. Through further combination optimization, four food-derived small molecule compounds with synergistic effects have been obtained. This serum-free culture medium can significantly promote the proliferation of bovine muscle stem cells, effectively maintain cell proliferation activity, and cells passaged using this serum-free culture medium can still maintain good differentiation potential. This provides a stable, sufficient, and safe cell source for the industrial production of cultured meat, which is beneficial for the large-scale production of bovine muscle stem cells.

A substrate for culturing cells that comprises a modified prolamin having a net positive charge is provided. The modified prolamin may be a prolamin that has been modified through amidation or esterification, such as the addition of a methyl-ester group or an ethanolamine group. The modified prolamin is animal-free may be food-grade and / or may be free of cross-linkers. The modified prolamin may be in a coating that can be applied to a surface for culturing adherent cells, such as a microcarrier (e.g., beads) or well plates. Substrates coated with the modified prolamin are able to support cell expansion and can be used for food-grade applications such as cultured meat.

The invention belongs to the technical field of cell culture meat screening, and particularly relates to a cell culture meat food hazard factor screening system and device.The cell culture meat food hazard factor screening system comprises a sensing unit, an execution unit, an analysis unit and a control unit; the sensing unit comprises an integrated sensor and living cell imaging equipment, the integrated sensor is used for collecting the culture environment of cell culture meat in real time, and the living cell imaging equipment is used for dynamically tracking the cell growth state; the execution unit is used for executing sample data acquisition and detection according to presetting; and the analysis unit analyzes and processes the data information sent by the sensing unit and the execution unit based on an AI data analysis platform. The intelligent level of screening the harmful factors of the cell culture meat can be greatly improved, so that dynamic evaluation and early warning of the harmful factors in the production process of the cell culture meat are realized, workers can timely and effectively perform corresponding treatment, and the production quality of the cell culture meat is further ensured.

The present invention aims to provide a cell culture substrate suitable for cell culture. In particular, it aims to provide a cell culture substrate that can efficiently produce cultured meat. [Solution] The cell culture substrate according to the present invention comprises a three-dimensional structure composed of proteins that facilitate cell adhesion and lipids that provide a hydrophobic environment conducive to cell adhesion. Therefore, animal cells can be cultured using a cell culture carrier equipped with this three-dimensional structure. Furthermore, in the cell culture substrate of the present invention, the sum of the dry mass of proteins and lipids in the dry mass of the cell culture substrate is 80% by mass or more, so animal cells adhere easily to the cell culture substrate, preventing unintended inhibition of culture. Based on the above, the cell culture substrate according to the present invention is a cell culture substrate suitable for cell culture.

The present invention relates to cultured animal cells having a genetic modification in the NF2 gene or having a modification to Merlin protein activity wherein the animal is an animal species suitable for human or animal consumption. Also provided are methods of producing cultured meat or cultured meat products 5 comprising culturing animal cells having such mutations.

The present invention relates to a composition for promoting differentiation of muscle stem cells into myotubes, comprising a compound that promotes lipid metabolism or cell differentiation, for example, ethanolamine or a functional equivalent thereof, and to a method for differentiating muscle stem cells using same. More specifically, the present invention provides a composition for promoting differentiation of muscle stem cells into myotubes, comprising only ethanolamine, a composition for a muscle stem cell culture medium comprising same, a method for differentiating muscle stem cells into myotubes, using the composition for promoting differentiation or the composition for a culture medium, a myotube culture obtained by the differentiation method, and a cultured meat and food composition comprising same.

The purpose of the present invention is to provide a culture medium in which cells to be used for producing a cultured meat can be grown, by adding thereto a food material as an additive. As a result

The invention discloses a co-culture method of cell culture meat and application of the co-culture method in improving meat quality and flavor. According to the invention, through two-stage differentiation, muscle precursor cells and fat precursor cells are respectively subjected to proliferation culture, then are separately subjected to primary differentiation, and are mixed together for co-culture, and meanwhile, second-stage continuous differentiation is carried out; the problem that differentiation of muscle cells and fat cells cannot be effectively and simultaneously differentiated under the co-culture condition due to different requirements of differentiation of muscle cells and fat cells on the serum environment is solved. In addition, the invention also provides an objective standard for evaluating the flavor quality of cell culture meat generated by co-culture. The cell culture meat with different flavors can be obtained by adjusting the proportion of the co-culture cells through the co-culture method, and the flavor quality of the cell culture meat from other sources can be evaluated and detected through the amino acid detection means.

Provided is a method for inducing muscle-derived differentiation of porcine pluripotent stem cells, which does not involve transgenosis and is free of serum addition in the whole course, and also provided is a method for preparing cell cultured meat (CM) based on the muscle-derived differentiation induction method.

Provided are: an improved culture medium composition which can be used for culturing pluripotent stem cells of mammals, preferably for establishing and maintaining pluripotent stem cells of mammals useful for producing cultured meat for cattle, pigs, or the like; and a culture method using the culture medium composition. The culture method according to the present invention comprises a step of culturing pluripotent stem cells of mammals in a culture medium composition, the culture medium composition being a culture medium composition for culturing pluripotent stem cells of mammals, the culture medium composition being a serum-free culture medium containing (1) a CDK8 / 19 inhibitor, (2) a ROCK inhibitor, (3) a SMAD activator, and (4) a STAT3 activator.

The present invention relates to the fields of biomaterials and food, and particularly to a method for physically crosslinking to prepare a cell-cultured meat biocompatible scaffold. The physical crosslinking method enables the internal combination of the biocompatible scaffold material through physical forces to form a biocompatible scaffold. The physical crosslinking method comprises the following steps: 1) shaping and drying the scaffold material; 2) steam crosslinking the dried scaffold material. The present invention uses a physical crosslinking method to prepare an edible protein or polysaccharide scaffold to support animal cell culture. No chemical crosslinking reagents are used during the production process, and no coatings such as ECM proteins are used. At the same time, cells proliferate well on the scaffold. The physical crosslinking step of this method can be combined with the sterilization step, with simple operating steps, low cost, and is applicable to the preparation of scaffolds of various forms.