119 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 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 of culturing cells using a food ingredient as an additive, it became possible to obtain high growth potential in fibroblasts and myoblasts by adding egg white or a dried product of egg white as a cell growth promoter. Accordingly, the present invention pertaining to a cell-growing culture medium containing a basic culture medium and egg white as a cell growth promoter was accomplished.

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.

The present invention belongs to the technical field of bioreactors, and in particular relates to a high-density culture method for porcine myoblasts based on a bioreactor. Through the steps of resuscitating porcine myoblasts, cell-microcarrier attachment treatment, bioreactor stirring mode conversion, counting and centrifugal cell collection, the porcine myoblasts are difficult to reach a high density in a bioreactor, and a harvest density of 10^6 cells / mL is achieved in the bioreactor, meeting the requirements of cell culture meat production on cell yield; at the same time, the culture time is shortened by more than 20%, greatly saving energy consumption cost and time cost.

The invention provides an application of dihydroflavonoid compound naringin in promoting proliferation and differentiation of porcine muscle stem cells to form myotubes, which is characterized in that the naringin is added into a DMEM / F12 culture medium containing 15% of fetal calf serum and 1% of double antibodies and a DMEM differential culture medium containing 2% of horse serum and 1% of double antibodies until the final concentration is 70 mu M, and the porcine muscle stem cells are respectively treated in proliferation and differentiation stages, hoechst 33342 staining and immunofluorescence staining methods are used for respectively judging the number of cells in the proliferation stage and the fluorescence area, fluorescence intensity and nuclear fusion rate of Myosin in myotubes formed in the differentiation stage. The relative proliferation multiple of the muscle stem cells treated by adding naringin and the fluorescence area, the fluorescence intensity and the nuclear fusion rate of Myosin in myotubes are obviously higher than those of a blank control group. By adopting the method, proliferation and differentiation of the porcine muscle stem cells can be promoted, and a new thought is provided for maintaining cell dryness and large-scale production of cultured meat.

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.

Edible microcarrier for growing anchorage-dependent cells to prepare a cultured meat product, where the edible microcarrier includes a microcarrier core and a microcarrier coating at least partially directly or indirectly covering the surface of said microcarrier core, wherein said microcarrier core is a hydrogel having only the following components: (A) 0.03-10.0 wt % of at least one ionically crosslinked polysaccharide; (B) 0.05-5 wt % of a mechanical stability agent different from (A); (C) 0.0-10 wt % of additives different from (A) and (B); (D) 0.0-50 wt % living cells; (E) 75-99.92 wt % of water; where the sum of (A)-(E) making up 100 wt %, and where the microcarrier coating includes a biopolymer, where the biopolymer is crosslinked using a crosslinking agent.

The present disclosure provides extruded or spun, semi-permeable, porous hollow fibres, comprising covalent ester, thioester and / or amide crosslinked polypeptides as well as processes for their production. The hollow fibres may be produced from protein, 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 fibres may be designed to be used in the production of cartridges that are compatible with existing and / or novel bioreactor platforms, for harbouring cell cultures in cultured meat production.

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 invention discloses a method for establishing a co-culture system of porcine skeletal muscle satellite cells (PSCs) and porcine precursor fat cells (ADSCs). The method comprises the following steps: selecting a PSCs and ADSCs co-culture proliferation culture medium and a co-culture differentiation culture medium, and establishing an optimal cell co-culture ratio. According to the optimal co-culture proliferation and differentiation culture medium for the PSCs and the ADSCs and the established co-culture proportion, mass proliferation and efficient differentiation of cells can be promoted, the influence of addition of fat cells with different concentrations on muscle cell differentiation can be researched, fat deposition in co-culture is regulated by regulating the proportion of the fat cells, and the differentiation of the muscle cells is promoted. And an earlier-stage foundation is provided for explaining a signal mechanism theoretical basis between cells under growth and development conditions of muscle and adipose tissues and exploring cell culture meat in a later stage.

The present disclosure provides extruded or spun, semi-permeable, porous hollow fibres, comprising covalent ester, thioester and / or amide crosslinked polypeptides as well as processes for their production. The hollow fibres may be produced from protein, 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 fibres may be designed to be used in the production of cartridges that are compatible with existing and / or novel bioreactor platforms, for harbouring cell cultures in cultured meat production.

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 invention relates to the field of bioengineering technology, and in particular to a single-cell green algae combined functional factor and its application in cell-cultured meat. The functional factor is obtained by subjecting the raw material to processes such as low-temperature freezing, room temperature thawing, and nano-scale shell adsorption column refining. The extraction process is simple and highly safe, and is suitable for industrial scale-up production. When applied to muscle cell culture, it can be directly dissolved in the basal culture medium, and the compounding method with the culture medium is simple. It has no toxic side effects on muscle cell growth and can completely replace serum for muscle cell culture. When the single-cell green algae combined functional factor mentioned in the present invention is applied to fish muscle cell culture, compared with the group without serum addition, it can provide cells with growth functional factor supplements, promote cell proliferation and differentiation in vitro, and can be applied to 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.

There is provided a substrate assembly for culturing cells, wherein the substrate assembly comprises: a plurality of edible fibres, wherein each fibre has an internal channel running along its length; a first support at a first end of the plurality of edible fibres; a second support at a second end of the plurality of edible fibres; wherein at least one of the first support and second support allows fluid communication across the support into the internal channels. Also provided is a method of culturing cells.