51 results about "Muscle stem cell" patented technology

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 serum-free culture medium for in-vitro proliferation of poultry muscle stem cells and application of the serum-free culture medium, and belongs to the technical field of cell engineering, a basic serum-free culture medium is DMEM-F12, a complete serum culture medium is a mixture of DMEM-F12 added with 10% of fetal calf serum and 1% of mycillin, and the volume ratio of a culture medium supplement to the basic culture medium is 1: 1000; the serum-free culture medium contains a supplement; the supplement comprises the following compositions: insulin, transferrin, fibroblast growth factor 2, epidermal growth factor, catalase, ethanolamine, vitamin B12, linoleic acid, linolenic acid, retinol acetate, tocopherol, L-alanine, L-proline, sodium selenite, sodium pyruvate, reduced glutathione, berberine hydrochloride and analogues thereof.

The invention discloses an application of NSUN2 in muscle injury repair. Research finds that the number of activated muscle stem cells of a CTX model mouse can be remarkably increased and proliferation of the muscle stem cells can be promoted by mediating NSUN2 overexpression through AAV, and the overexpression of the NSUN2 not only improves the activation level of the muscle stem cells, but also enhances the self-renewal capacity of the muscle stem cells; in addition, overexpression of NSUN2 also significantly increases the area of muscle fibers of CTX model mice and reduces the weight of muscle atrophy. Overexpression of the NSUN2 promotes muscle fiber repair and muscle mass recovery by promoting proliferation and activation of muscle stem cells, so that healing of muscle injury is promoted, and the NSUN2 has a treatment effect on the disease. Therefore, the invention provides a new method for the treatment of muscle injury, and a new way is possibly opened up for the treatment of other muscle-related diseases.

The present disclosure provides methods for culturing primary muscle stem cells, promoting self-renewal of primary muscle stem cells in cell culture, inhibiting the differentiation of primary muscle stem cells and increasing the clonogenicity of primary muscle stem cells, comprising culturing primary muscle stem cells in the presence of an inhibitor of mitochondrial maturation. Also provided are methods for promoting the differentiation of primary muscle stem cells, comprising culturing primary muscle stem cells in the presence of an promoter of mitochondrial maturation. The present disclosure also provides the use of inhibitors of mitochondrial maturation and promoters of mitochondrial maturation in such methods, and kits comprising the same.

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.

Disclosed herein are hydrogels that can be loaded with myogenic agents, for instance muscle stem cells (MuSCs) / satellite cells, pro-myogenic factors, and combinations thereof. The hydrogels can be contacted with damaged muscle tissue, thereby facilitating muscle growth and repair. Further provided are methods of repairing muscle tissue in a patient in need thereof, said methods comprising contacting the muscle tissue with a composition comprising the hydrogel. Also provided herein are kits comprising the hydrogel composition and a substrate comprising at least one microneedle.

This invention discloses the application of the Six2 gene in promoting the reprogramming of fibroblasts into muscle stem cells, belonging to the field of biotechnology. The nucleotide sequence of the Six2 gene is shown in SEQ ID NO.1, and the fibroblasts are chicken fibroblasts. This invention uses chicken embryonic fibroblasts as the reprogramming target, and induces the reprogramming of chicken embryonic fibroblasts into muscle stem cells by overexpressing the Six2 gene, promoting the expression of stem cell-related genes MyoD, MyoG, Myf5, and Pax7. The circ_TCONS / gga-miR181-5p / Six2 regulatory network participates in the reprogramming process; it improves the efficiency of somatic cell reprogramming into stem cells, greatly promotes the application of reprogramming, and provides a new approach for the sustainable acquisition of muscle stem cells.

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 application discloses a kind of construction method and application of seawater fish muscle stem cell line, culture medium with basic medium as background medium;Culture medium includes LIF growth factor, EGF growth factor and leopard bristletooth-derived FGF2 protein;The final concentration of leopard bristletooth-derived FGF2 protein is 20~30 ng / mL, 30~45 ng / mL, 45~55 ng / mL or 55~70 ng / mL. Through the medium, immortalized seawater fish muscle stem cell line with good stability, cell division vigorous, short subculture time can be constructed, and can be stably subcultured for more than 50 generations. It provides raw materials for exploring the function of myoblasts, gene regulation mechanism, and provides the prerequisite for in vitro verification experiment for further exploring the influence of seawater fish muscle stem cells on growth rate, and lays a foundation for good strain breeding of seawater fish.

The invention relates to a living plant cell-animal cell composite scaffold as well as a preparation method and application thereof. The living plant cell-animal cell composite scaffold comprises a 3D printing hydrogel scaffold used as a muscle repair carrier and a 3D printing biological ceramic scaffold used as a bone repair carrier, wherein the 3D printing hydrogel scaffold and the 3D printing biological ceramic scaffold are assembled by simulating a muscle-skeleton structure; the hydrogel scaffold comprises a polymer matrix component, and functional plant cell components, namely diatom cells and mammalian cell components, namely muscle stem cells, which are distributed in the polymer matrix component, the biological ceramic scaffold comprises a biological ceramic scaffold matrix component and mammalian cell component bone marrow mesenchymal stem cells inoculated and loaded on the biological ceramic scaffold matrix component.

The application discloses application of a polysaccharide wrapping film in preparation of a cell culture product and belongs to the technical field of cell culture food. The application specifically forms the polysaccharide wrapping film on the periphery of a cell inoculation support after the cell inoculation support, can relieve the phenomenon that cells fall off from the support during culture, and further guarantees effective proliferation and differentiation of muscle stem cells and / or fat stem cells on the support, thereby providing a new solution approach for efficient and standardized production of the cell culture product.

The present invention provides: a serum replacement composition for cell culture, comprising an extract of Chlorella protothecoides; and a method for culturing cells, such as muscle stem cells, using the composition.

This application discloses a muscle stem cell resuscitation and protection washing solution, a quality assessment kit, and a method of use, belonging to the field of cell biology technology. In this application, vitamin E succinate and mannan peptide are used as core antioxidant components, synergistically combined with Gynostemma pentaphyllum polysaccharide, Portulaca oleracea polysaccharide, trehalose, L-proline, and magnesium sulfate heptahydrate to construct a four-in-one resuscitation and protection washing solution that provides isotonic buffering, extracellular protection, intracellular protein stabilization, antioxidant effects, and mitochondrial stabilization. This increases the survival rate of muscle stem cells after resuscitation to over 93%. The kit integrates a multi-dimensional detection system for cell resuscitation and protection washing solution, cell viability, proliferation rate, Pax7 / MyoD double-positive stemness assessment, and cryopreservation damage, achieving closed-loop management of the entire process of muscle stem cell resuscitation, protection, and quality assessment. The double-positive rate of stemness markers reaches over 90%, providing reliable quality assurance for cell bank management and clinical applications.

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.

This invention discloses a method for constructing and applying a marine fish muscle stem cell line. The culture medium uses basal medium as the background medium and contains LIF growth factor, EGF growth factor, and FGF2 protein derived from the leopard-gill sea bass. The final concentration of FGF2 protein from the leopard-gill sea bass is 20-30 ng / mL, 30-45 ng / mL, 45-55 ng / mL, or 55-70 ng / mL. Passaging using this medium can construct a stable, rapidly dividing, and short-passaging immortalized marine fish muscle stem cell line, capable of stable passage for more than 50 generations. This provides raw materials for exploring the function and gene regulation mechanisms of myoblasts, and provides in vitro verification experimental conditions for further research on the effects of marine fish muscle stem cells on growth rate, laying the foundation for the breeding of superior marine fish species.

Disclosed are methods of in situ genomic modification of cells (e.g., stem cells, tissue stem cells, muscle stem cells, Sca-1+ mesenchymal progenitor cells in skeletal muscle, CD140a+ dermal mesenchymal cells) using sequence-targeting nucleases delivered via a virus (e.g., an AAV).

Disclosed are methods of in situ genomic modification of cells (e.g., stem cells, tissue stem cells, muscle stem cells, Sca-1+ mesenchymal progenitor cells in skeletal muscle, CD140a+ dermal mesenchymal cells) using sequence-targeting nucleases delivered via a virus (e.g., an AAV).

The application provides a skeletal muscle organoid and a preparation method thereof, takes muscle stem cells, neural stem cells and fibroblasts as primary cells for organoid culture, restores the microenvironment of skeletal muscle as much as possible, and improves the differentiation degree of the organoid; further, the mold and frame for constructing the organoid are improved, the batch production of the organoid is realized through the array design of the PDMS mold and the polyamide frame; finally, the calcium signal video of the organoid is analyzed by using the Organblink system independently developed by the application, and the accuracy of the function evaluation result of the organoid is improved. The method provided by the application realizes the high bionics and high batch production of the skeletal muscle organoid, has low cost, and greatly promotes the development of the medical research field.

A composition of INFRA cells is described. The composition includes human muscle stem cells (SCs) expressing the transcription factor PAX7 and lacking the expression of the transmembrane receptor PDGFRA, wherein at least 70% of the population of human muscle SCs are PAX7+ and PDGFRA-. Methods of preparing the INFRA cell compositions, and methods of using the INFRA cells to treat muscle injury are also described.

The invention discloses application of a Six2 gene in promoting reprogramming of fibroblasts into muscle stem cells, and belongs to the technical field of biology. The nucleotide sequence of the Six2 gene is as shown in SEQ ID NO.1, and the fibroblast is a chicken fibroblast. Chicken embryo fibroblasts are adopted as reprogramming objects, the chicken embryo fibroblasts are induced to be reprogrammed into muscle stem cells in a mode of overexpression of Six2 genes, expression of stem cell related genes MyoD, MyoG, Myf5 and Pax7 is promoted, and a circTCONS / gga-miR181-5p / Six2 regulatory network participates in the reprogramming process; the efficiency of reprogramming the somatic cells into the stem cells is improved, the application of reprogramming is greatly promoted, and a new way is provided for continuously obtaining the muscle stem cells.

The present disclosure provides, inter alia, compositions and methods for treating premature aging of muscle stem cells to combat myopathy by targeting Cdkn2a.

The invention belongs to the technical field of exosome biological medicines, and particularly relates to a preparation method and application of a drug-loaded muscle fiber exosome for improving sarcopenia of a pet dog. The method is realized by the following steps: firstly, separating and culturing high-activity muscle stem cells: culturing and amplifying the high-activity muscle stem cells; then culturing, amplifying and differentiating the cultured primary muscle stem cells (MPCs) to form muscle fiber cells; the differentiated muscle fiber cells are loaded with SR12343 in several times, and the drug-loaded muscle fiber exosome is obtained. The muscle fiber exosome prepared by the invention contains specific bioactive factors of muscle functional cells, and can effectively improve the activity of an NF-kB signal channel for inhibiting mediating cell senescence and promote the activation and regeneration of senescent muscle cells in vivo under the action of cooperating with SR12343.

Provided is a method for loading a muscle stem cell exosome with a Notch activator and a senolytic drug, fisetin, that is capable of targeting and killing senescent cells, and further with lutein that has an eye protection effect, such that senescent muscle cells can be eliminated while the regeneration of muscle stem cells is promoted. By means of high-activity stem cell separation and culture technology, combined with exosome large-scale preparation and separation technology, simple and low-cost exosome drug-loading technology, senescent cell targeted clearance technology, etc., the present invention effectively helps to ameliorate the problems of extraocular muscle aging and atrophy, and can be widely applied to the damage repair of various muscle tissues. The present invention provides a new option for ameliorating the symptom of degeneration of extraocular muscle function, and has outstanding advantages in terms of safety, biocompatibility, and effectiveness.

The application discloses a primer set for real-time fluorescent quantitative PCR detection of a pig muscle regulatory factor and application thereof, and belongs to the technical field of biology. The application provides specific primer pairs capable of detecting pig muscle regulatory factors MyoD and MyoG and reference primer pairs. cDNA is obtained by extracting sample RNA and performing reverse transcription, and real-time fluorescent PCR is performed by using the provided primers, so that the mRNA expression amount of the pig muscle regulatory factors MyoD and MyoG can be simultaneously detected quickly, specifically, sensitively and stably, and the application can be used for detecting the change in the expression level of the regulatory factors before and after the differentiation of pig muscle stem cells, so that the differentiation condition of pig breeding meat can be judged.

This invention relates to the field of bioengineering technology, specifically to an extracellular vesicle from kelp, its preparation method, and its applications. This invention provides a method for efficiently isolating extracellular vesicles with specific biological activities from kelp, and an application scheme for these vesicles to promote the proliferation, migration, and maintenance of stemness of large yellow croaker muscle stem cells under low serum conditions. This overcomes the problems of high cost and limited functionality of fetal bovine serum as a substitute in existing cell-cultured meat technologies, providing a novel culture medium supplement component that is stable in origin, low in cost, and has a clear effect.

A composition contains Nicotinamide and Vitamin B6, for example pyridoxine. The composition may be an oral nutritional composition, for example an infant formula, a follow up formula, a nutritional supplement, an oral nutritional supplement, a food product. The composition can be administered to an individual in need thereof for promoting muscle growth and / or for maintaining and / or increasing muscle function and / or muscle mass in infants and children. For example, the composition can be useful for increasing muscle function and / or muscle mass by modulating muscle stem cells.

The invention discloses a porcine serum culture medium and application thereof in porcine muscle stem cell culture, and belongs to the technical field of animal cell culture. The porcine serum culture medium comprises a basic culture medium, porcine serum and additive factors, wherein the volume percentage of the pig serum is 10-20%, and the additive factor comprises at least one of the following components: a chick embryo culture with the volume percentage of 0.3-0.7%, glutamine with the volume percentage of 0.8-1.2%, non-essential amino acid with the volume percentage of 0.8-1.2%, and a fibroblast growth factor-2 with the mass concentration of 2.3-2.7 ng / mL. According to the porcine serum culture medium provided by the invention, by adding the porcine serum, the cost can be remarkably reduced, and meanwhile, the multiplication capacity of cells can also be improved; in addition, cell proliferation can be further promoted by adding additive factors; therefore, the porcine serum culture medium has a relatively good application prospect in culture of porcine muscle stem cells.

The invention discloses an application of PDGF-C (Platelet Derived Growth Factor-C) in muscle injury repair. Research finds that in a CTX model mouse with Pdgf-c knocked out, the number of proliferated muscle satellite cells (Pax7 + Ki67 +) is remarkably smaller than that of wild type mice. Through AAV-mediated PDGF-C overexpression, the number of CTX model mouse muscle stem cells can be significantly increased, the activation level of the muscle stem cells is improved, and the self-renewal ability of the muscle stem cells is enhanced. In addition, PDGF-C overexpression also significantly increases the area of muscle fibers of CTX model mice and improves the regeneration and repair ability of the muscle fibers, thereby promoting the healing of muscle injury and having a therapeutic effect on the disease. In addition, the effect of the PDGF-C on promoting muscle injury repair is obviously higher than that of the PDGF-B. Therefore, the invention provides a new method for the treatment of muscle injury, and a new way is possibly opened up for the treatment of other muscle-related diseases.

This invention discloses the application of PDGF-C in muscle injury repair. The study found that in CTX model mice with PDGF-C knockout, the proliferation of muscle satellite cells (Pax7)... + Ki67 + The number of muscle stem cells in CTX model mice was significantly lower than that in wild-type mice. AAV-mediated PDGF-C overexpression significantly increased the number of muscle stem cells, improved their activation level, and enhanced their self-renewal capacity. Furthermore, PDGF-C overexpression significantly increased the surface area of ​​muscle fibers in CTX model mice, enhancing their regenerative and repair capabilities, thereby promoting muscle injury healing and demonstrating a therapeutic effect on this disease. Moreover, PDGF-C's effect on promoting muscle injury repair was significantly higher than that of PDGF-B. Therefore, this invention provides a new method for treating muscle injuries and may open new avenues for the treatment of other muscle-related diseases.

The invention provides a low-cost, safe and serum-free culture medium for in-vitro efficient suspension culture of porcine muscle stem cells and application of the serum-free culture medium. Comprising cell culture supplementary factors and a basic culture medium, the cell culture supplementary factors comprise zinc sulfate, L-ascorbic acid, bFGF, IGF-1, LIF, EGF, cortisol, HGF, alpha-tocopherol, PVA, beta-cyclodextrin, methyl cellulose and a yeast extract. The efficient suspension amplification of the porcine muscle stem cells under serum-free and carrier-free conditions is realized by adding factors in a simplified and optimized manner and adding cell culture supplementary factors with specific types and concentrations. According to the culture medium disclosed by the invention, the addition cost of exogenous factors is remarkably reduced, various potential risk substances are avoided, the food safety and economy of the culture medium are improved while efficient growth of cells is ensured and the process is simplified, and an important technical and raw material basis is provided for industrial production of cell culture meat.