62 results about "Muscle stem cell" patented technology

Compositions and methods for modulating proliferation and / or lineage commitment of stem cells by modulating the Wnt signalling pathways. Modulators of the Wnt signalling pathways and screening methods to identify modulators are also provided. The methods of the invention may be conducted in vitro or in vivo to induce or inhibit stem cell proliferation and / or lineage commitment, and are particularly useful for in vivo stimulation of proliferation and / or lineage commitment of resident stem cells in a tissue.

Disclosed are methods of differentiating stem cells into muscle cells by growing the cells in a myogenic culture medium. The differentiated cells can be used as a source of cells for transplantation in a patient in need thereof.

The invention discloses an in-vitro efficient culture method for muscle stem cells, and a universal muscle stem cell in-vitro culture kit comprising a muscular tissue digestive enzyme, a muscle cell culture medium and a cell culture plate coating solution. The in-vitro culture method is characterized by being capable of rapidly acquiring and amplifying muscle stem cells, maintaining regeneration activity of the muscle stem cells and culturing stem cells which are stable in cell morphology, capable of stably expressing a muscle stem cell marker Pax7 and high in proliferation activity. By adopting the technique disclosed by the invention, relatively pure mouse muscle stem cells can be obtained, and the muscle stem cell marker Pax7 can be expressed. In addition, the cells also express muscle system transcription factors Myf-5 and MyoD; after the cells are differentiated into myotubes, a distinctive muscle structure protein Desmin can be stably expressed.

The present invention relates to the use of antibodies recognizing Fetal Antigen-1 (FA1 / dlk1) for the detection and isolation of cell populations in mammalian muscle. In one embodiment, myogenic progenitor cells are detected in developing, diseased or regenerating muscle. In another embodiment, muscle stem and progenitor myogenic progenitor cells are isolated from muscle tissue or from cultures containing muscle cells. The isolated cells may be used for transplantation, drug screening, production of cell type specific antibodies, and gene therapy and discovery. Transplantation of these cells may provide treatments for degenerative diseases of muscle, and for regeneration of muscle following trauma or ischemia such as myocardial infarction.

The invention discloses a preparation method and application of cross-linked hydrogel for muscle stem cell culture, and belongs to the technical field of biological food materials. Chitosan, alginate, dextran and Ca<2+> are crosslinked through a physical crosslinking method to form double-network hydrogel with high mechanical strength, and then the hydrogel is coated with heparin and collagen through a dipping coating method, so that the hydrogel can immobilize growth factors and adhere to cells. Meanwhile, the extracted primary muscle stem cells are inoculated onto the hydrogel and cultured for 24 hours in a growth culture medium (79% of DMEM, 10% of FBS and 1% of double antibodies); and then the extracted primary muscle stem cells are cultured in an incubator with a differential medium (97% of DMEM, 2% of horse serum and 1% of double antibodies) for 7 days. By utilizing the hydrogel, the absorption of the muscle stem cells on nutrient substances can be enhanced, and the growth of the muscle stem cells is facilitated. The dual-network hydrogel has the potential to be used as a scaffold for muscle stem cell growth of stem cell culture meat.

The invention discloses a method for promoting proliferation of muscle stem cells, and belongs to the technical field of cell culture and biology. The invention designs a culture medium for effectively amplifying muscle stem cells in vitro. The culture medium comprises L-ascorbic acid, D-ascorbic acid and / or insulin-like growth factor-I, After the muscle stem cells cultured by the optimized culture medium are cultured for 21 days, the total cell number is increased by more than 10<6> times, and the proportion of Pax7 <+> cells still exceeds 60%; the amplified muscle stem cells can form a largenumber of typical muscle tubes through induced differentiation, and express muscle fiber specific proteins MyHC and Actin. and effective amplification of the muscle stem cells is achieved, and the method has important significance in large-scale culture of the muscle stem cells for preparing stem cell grafts and producing cell culture meat.

The invention relates to the technical field of medical and health-care products and particularly relates to a method for preparing a stem-cell active-factor composition with a muscle cell repairing function. The method comprises the steps: firstly, acquiring autogenous or xenogenous human muscle stem cells with active regeneration capacity, and carrying out mass amplification through in-vitro culture, so as to establish a cell line for long-term application; and breaking the cells with ultrasonic waves under low-temperature conditions, carrying out centrifugation, then, collecting supernatant, carrying out filtering and concentrating so as to obtain a muscle stem cell extract, merging an extract of a stem cell culture solution, adding an activity stabilizer and a preservative, and then, carrying out mixing with a penetrant, thereby obtaining the active-factor composition. The method has the beneficial effects that the stem-cell active-factor composition can exert very good repairing and regrowing actions on impaired muscle and skin; and an improvement on a senescence state of the skin can be promoted.

The object of the present invention is to provide a method for screening a substance involved in a metabolic shift of skeletal muscle, and a kit for screening a substance involved in a metabolic shift of skeletal muscle.The object can be solved by a muscle stem cell or myoblast comprising at least one myosin-heavy chain fusion gene selected from the group consisting of a myosin-heavy chain I fusion gene wherein a myosin-heavy chain I gene and a fluorescent protein or photoprotein gene are fused, a myosin-heavy chain IIa fusion gene wherein a myosin-heavy chain IIa gene and a fluorescent protein or photoprotein gene are fused, a myosin-heavy chain IId / x fusion gene wherein a myosin-heavy chain IId / x gene and a fluorescent protein or photoprotein gene are fused, and a myosin-heavy chain IIb fusion gene wherein a myosin-heavy chain IIb gene and a fluorescent protein or photoprotein gene are fused.

The invention relates to application of Linc-RAM for treating muscle diseases. Specifically, the invention relates to application of Linc-RAM in preparation of medicines for treating muscle diseases, particularly muscle injury and rhabdomyosarcoma. Furthermore, the invention relates to application of the Linc-RAM in preparation of medicines for promoting differentiation of muscle stem cells, as well as reagents and medicines for promoting transition of fibroblast to myoblast.

The invention provides an improved culture medium for in-vitro amplification culture of cultured meat seed cells, namely currently optimal seed cells (muscle stem cells) of cultured meat. The improvedculture medium is a cell culture medium added with VC and derivatives thereof, cells with the number more than 1000 times that of a normal culture medium can be obtained in in-vitro amplification culture, and more importantly, the muscle stem cells are subjected to in-vitro long-term continuous passage and can still keep differentiation potential after the required cell number is reached. Therefore, the improved culture medium can help mass amplification culture of functional muscle stem cells in vitro, and better meets the large-scale requirements of cultured meat production on seed cells.

The invention discloses cross-linked hydrogel for muscle stem cell culture as well as a preparation method and application of the cross-linked hydrogel, and belongs to the technical field of biological food materials. The preparation method comprises the following steps: dissolving collagen to prepare a solution, then adding a certain amount of alginate and heparan sulfate proteoglycan, and uniformly mixing the alginate and heparan sulfate proteoglycan with the collagen solution; and adding epsilon-PL and TG enzyme into the solution, uniformly conducting stirring, and putting slurry into a mold for crosslinking to obtain the hydrogel. According to the hydrogel disclosed by the invention, collagen, polylysine and heparan sulfate proteoglycan are connected through TG enzyme to form covalent crosslinking, and a compact three-dimensional 'egg box' net structure is formed through physical electrostatic interaction between polylysine and alginate. By utilizing the hydrogel, the absorption of the muscle stem cells on nutrient substances can be enhanced, and the growth of the muscle stem cells is facilitated. The bi-crosslinked network hydrogel has the potential to be used as a scaffold for muscle stem cell growth of stem cell culture meat.

The invention provides a bioactive substance composition, a serum-free culture medium containing the composition and application of the serum-free culture medium. The bioactive substance composition is used for a serum-free culture medium and / or a composition and preparation thereof; the serum-free culture medium and / or composition can be used for primary culture and subculture of cells and / or tissues. The cells are selected from any one or more of tendon and / or ligament source cells, chondrocytes, meniscus stem cells, mesenchymal stem cells, skeletal stem cells and muscle stem cells. The tissue is a source tissue of a motor system. The bioactive substance composition and / or the serum-free culture medium and / or the composition can be used for preparing a medicine for treating tissue and / or organ injury; the tissue or organ injury is selected from movement system tissue or organ injury.

Provided is a muscle stem cell in vitro culture method. A muscle stem cell is cultivated in vitro by using a cell culture medium of a cell factor added with a blood cell or a conditioned medium of the blood cell. Also provided is a culture medium used in the muscle stem cell in vitro culture method and an application thereof.

The invention belongs to the field of stem cell application, and particularly relates to a method for preparing animal cell culture meat from high-activity muscle stem cells. A polypeptide fragment of Jagged1 is combined in the hydrogel cell scaffold in advance, and a Notch signal channel of the stem cells is effectively activated, so that the characteristics (dryness and growth differentiation potential) of the stem cells are maintained. The added Follistatin can inhibit the activity of myostatin, so that the volume, growth and differentiation of muscle cells are more potential, and the differentiated muscle fibers are thicker and stronger. And the more mature and robust muscle fibers can enhance the texture and taste of the cell culture meat and generate more special nutritional factors of the mature muscle cells. Meanwhile, the hydrogel thin layers on which the muscle cells grow are overlaid in multiple layers to form the three-dimensional muscle tissue, and differentiation of cell myogenic fibers under the low-serum condition is continued, so that the growth and differentiation time and space advantages of the muscle stem cells are arranged more reasonably.

The invention discloses an application of muscle stem cells in preparation of an anti-inflammatory drug. The muscle stem cells or the supernate thereof are mediated by anti-inflammatory macrophages, the anti-inflammatory characteristic is formed in the macrophage maturation process, and inflammatory diseases caused by inflammatory macrophages as main pathogenic cell populations can be effectivelytreated. For example, the disease symptoms of weight loss, diarrhea, hematochezia, intestinal tissue thickening and edema, structural damage and immune cell infiltration can be effectively relieved bytreating colitis mice with muscle stem cells or supernate thereof.

Disclosed herein are agents that enhance muscle stem cell engraftment, as well as methods and compositions using the same.

An in vitro method of producing a population of muscle stem cells comprising co-culturing pluripotent stem cells, embryonic fibroblast cells and endothelial cells in 3D cell culture.

The invention discloses a method for separating sheep muscle stem cells. According to the method, muscle cells of the legs of a sheep are cut and obtained, the muscle cells are disinfected, cleaned, chopped, digested, centrifugally separated and the like, the muscle stem cells are separated and placed in a petri dish for culture; compared with conventional methods for separating the muscle stem cells, the method has the advantages that the separation efficiency is high, the infection rate in the separation process is low, the separated stem cells are high in survival rate and easy to culture,the number of the stem cells is increased, the muscle repair speed in the process of using the stem cells for treatment is increased, and the muscle repair efficiency in the process of using the stemcells for treatment is improved.

The invention provides a chemically defined medium for in vitro differentiation of muscle stem cells, that is, a serum-free, more efficient and cheap chemically defined medium for inducing differentiation of muscle stem cells in vitro. Compared with the existing general muscle stem cell differentiation medium, this chemically defined medium can increase the relative expression of MYOG gene by 4.48 times on the second day of differentiation, and increase the relative expression of MYHC gene by 55.28 times on the sixth day of differentiation , the percentage of cell differentiation at the terminal differentiation stage increased from 34.94% to 57.93%, there was a very significant difference, and the induced differentiation formed more, thicker and longer muscle fibers. The invention of the chemically defined differentiation medium further improves the differentiation efficiency of muscle stem cells, provides a more efficient and inexpensive method for muscle stem cells to differentiate into myotubes, and for 3D culture of muscle stem cells to produce cell cultured meat.

The invention provides an improved culture medium for short-term proliferation of cultured meat seed cells, which can accelerate the proliferation speed of the current most potential seed cells-muscle stem cells of cultured meat in short-term culture in vitro. The improved culture medium is a cell proliferation culture medium added with water-soluble vitamin E, the proliferation speed of the cells cultured by the improved proliferation culture medium in short-term culture in vitro is significantly improved, and the number of the cells is 1.2 times of that of the cells cultured by a normal culture medium; and the cells cultured by the improved proliferation culture medium added with the water-soluble vitamin E are induced and differentiated in a differentiation culture medium, so that the differentiation ability of the cells can be significantly improved, and more muscle proteins can be produced. Therefore, the improved proliferation culture medium can help to obtain a large number of cultured meat seed cells in short term in vitro, and plays a very beneficial role in the production of the cultured meat.

The invention provides an application of muscle stem cells to preventing, relieving and treating metabolic disorder diseases. Researches find that the muscle stem cells can significantly improve the glycometabolism ability of the body, effectively control the blood glucose level, reduce white adipose tissue in the body, relieve the swelling and increase of adipose cells, improve the steady state of the white adipose tissue, reduce the inflammatory environment of the body, reduce the lesion degree of fatty liver and the like. In addition, the invention also provides a prevention and treatment method of muscle stem cells for increasing muscle volume and strength.

The invention provides a method of using umbilical cord blood stem cells to treat muscular dystrophy, and relates to the technical field of medical technology. The method comprises the steps of obtaining HLA fitted umbilical cord blood; based on the immunomagnetic bead method, enriching CD34+ cells; after myeloablative treatment, transplanting 50% of the CD34+ cells and all CD34- cells into a patient's body; increasing the conversion rate of the remaining 50% of the CD34+ cells through factor combination to transform the umbilical cord blood stem cells into muscle stem cells; and transplanting the middle stem cells to the patient. The transplanted muscle stem cells differentiate into functional muscle stem cells, and the muscle stem cells propagate and differentiate into muscle tissues. The hemopoietic stem cells can be converted into muscle stem cells, and the muscle stem cells originating from the differentiation of umbilical cord blood stem cells are transplanted, and therefore the method has a good transplantation capability and can improve or cure myotrophy sickness.

Disclosed are methods of genomic modification of skeletal and cardiac muscle using sequence-targeting nucleases and a donor sequence delivered via a virus.

The invention discloses a method for low-cost in-vitro culture of porcine muscle stem cells, and belongs to the technical field of stem cell and animal cell culture. The invention discloses a culturemedium and the culture method for culturing the muscle stem cells at low cost, the muscle stem cells can keep in-vitro growth and myogenic differentiation capacity through the culture medium and the culture method, and the cost of the culture medium is greatly reduced.

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 invention belongs to the technical field of cell engineering, and particularly relates to an extraction method for separating muscle tissues of bovine muscle stem cells and application of the extraction method. The method comprises the following steps: (1) wiping the surface of cattle skin with alcohol, collecting muscular tissues of longissimus on the back of the cattle, and immediately rinsing with a phosphate buffer solution; (2) removing fascia and white film on the surface; and (3) rinsing with triple distilled water, and then rinsing with a phosphate buffer solution, alcohol and a phosphate buffer solution in sequence. The extraction method is different from the conventional method for treating by singly using a phosphate buffer solution, and the cost is reduced by combined treatment of three washing solutions. The method comprises the following steps: extracting muscle tissue, separating muscle stem cells from the extracted muscle tissue, using the muscle stem cells to construct a super enhancer, and optimizing and forming a method for analyzing the bovine muscle super enhancer by using an H3K4me1 and H3K27ac double-labeling method on the basis of chromosome co-immunoprecipitation high-throughput data basic analysis, which is different from a single labeling method in the prior art.

The invention relates to programmed necrosis of muscle fiber cells after muscle tissue injury and a proliferation mechanism of muscle stem cells and application of the programmed necrosis and the proliferation mechanism. Muscle fibers with programmed necrosis highly express and release TNC, and the TNC promotes rapid proliferation of muscle stem cells by combining and activating EGFR on the surfaces of the muscle stem cells, so that damaged muscle tissues are repaired. The invention also provides a composition containing TNC, which can promote in-vitro mass amplification and long-term passage of muscle stem cells.