42 results about "C2C12" patented technology

The present invention provides methods of improving the osteogenic and / or chondrogenic activity of a bone matrix, e.g., a dermineralized bone matrix (DBM), by exposing the bone matrix to one or more treatments or conditions. In preferred embodiments the bone matrix is derived from human bone. The treatment or condition may alter the structure of the bone matrix and / or cleave one or more specific proteins. Cleavage may generate peptides or protein fragments that have osteoinductive, osteogenic, or chondrogenic activity. Preferred treatments include collagenase and various other proteases. The invention further provides improved bone and cartilage matrix compositions that have been prepared according to the inventive methods and methods of treatment using the compositions. The invention further provides methods of preparing, testing, and using the improved bone matrix compositions. On a assay comprises exposing relatively undifferentiated mesenchymal cells to a bone matrix composition and measuring expression of a marker characteristic of osteoblast or chondrocyte lineage(s). Increased expression of the marker relative to the level of the marker in cells that have been exposed to a control matrix (e.g., an inactivated or untreated matrix) indicates that the treatment or condition increased the osteogenic and / or chondrogenic activity of the bone matrix. Suitable cells include C2C12 cells. A suitable marker is alkaline phosphatase. The inventive methods increase the osteogenic and / or chondrogenic activity of human DBM when tested using this assay system.

The method of the invention provides for producing a heterologous protein in mammalian host cells having nucleic acid encoding Hepatitis B X protein and the heterologous protein, by growing mammalian host cells selected from the group consisting of HKB11, CHO, BHK21, C2C12, and HEK293 cells, by growing mammalian host cells in non-adherent suspension culture, or by growing mammalian host cells which contain nucleic acid providing exogenous X-box Binding Protein, XBP1s. The conditions should be such that HBx, exogenous XBP1s if present, and the heterologous protein are expressed by the mammalian cells. The invention includes compositions for carrying out the method.

The invention provides a method for promoting GLUT4 gene expression in a muscle cell, relates to a method for promoting GLUT4 gene expression, aims to build a method for effectively improving GLUT4 gene expression, and particularly relates to a method for promoting GLUT4 gene expression in the muscle cell to promote glucose absorption of the muscle cell. The method comprises the following steps: firstly, performing induction and differentiation culture on a muscle precursor cell (C2C12) so as to enable the muscle precursor cell to be cultivated to a mature muscle cell, then adding 4-PBA with the concentration of 1-10 mol / L into a culture medium, culturing for 12-24 h under the condition that the temperature is 37 DEG C and the volume concentration of CO2 is 5%, and recycling the cell; then detecting GLUT4 gene expression of the muscle cell and glucose absorption change of the muscle cell. The method provided by the invention is applied to the field of scientific research and clinical application for reducing diabetes hyperglycemia.

The invention discloses a bionic skeletal muscle composite tissue prepared by multi-channel extrusion 3D bioprinting. The preparation method comprises the following steps: preparing bone scaffold bionic bioink, periosteum bionic bioink, myofibrillar membrane bionic bioink, and muscle bionic bioink; Mix MSCs and C2C12 with the corresponding bionic bio-ink; use a multi-channel extrusion 3D bioprinter to print and form a four-layer composite tissue engineering scaffold of bionic bone, bionic periosteum, bionic sarcolemma, and bionic muscle. The bionic skeletal muscle composite tissue prepared by multi-channel extrusion 3D bioprinting of the present invention can minimize fibrosis during the recovery of traumatic skeletal muscle injury; the bionic skeletal muscle composite tissue prepared by multi-channel extrusion 3D bioprinting can simultaneously replace The structure and function of bone and skeletal muscle, supporting the proliferation and differentiation of myoblasts and osteoblasts; and using 3D bioprinting technology to make implants easy to customize to adapt to any defect shape.

The present invention belongs to the field of pig molecule genetics, and specifically relates to cloning identification of a pig muscle-specific expression gene integrin beta 1 binding protein 2 (ITGB1BP2) promoter, and an application thereof. The steps of the present invention comprise: extracting total RNA of different tissues of a large white pig, and carrying out fluorescence quantitative PCR to determine specific expression in muscle tissue. According to the present invention, the total DNA of the muscle tissue of the large white pig is adopted as a template to design primers; PCR amplification is performed to obtain a 1442 bp sequence of flank on 5' terminal of pig ITGB1BP2 gene; luciferase reporter gene plasmids of 15 ITGB1BP2 promoter deletion fragments with different lengths are constructed; and C2C12 cells are transfected with the plasmids, activity of the promoter of each deletion fragment is detected, and the pig ITGB1BP2 gene promoter is identified. The nucleotide sequence of the pig ITGB1BP2 gene promoter of the present invention is represented by SEQ ID NO:1. With the present invention, import elements and means are provided for pig genetic improvement and transgene, and muscle tissue specificity and stability of transgene expression can be strengthened.

The invention belongs to differentiation culture of myoblasts, and relates to a differentiation culture method of C2C12 myoblasts. Separated mouse myoblasts sub-cultured more than five times are used as objects, a growth medium and a differentiation medium are optimized through steps of cell proliferation culture, differentiation culture and the like, and over 85% of the myoblasts sub-cultured more than five times can differentiate myotubes, thus effectively improving the utilization ratio of the separated primary myoblasts.

The invention provides a solution for preserving magnetosome and an application thereof and belongs to the application field of biomaterials. The solution for preserving magnetosome is a phosphatic buffer solution (PBS). During the process of preserving magnetosome, the structure and functions of magnetosome are the least influenced and interfered by the solution. A BMPs-PEI / DNA composite obtained by connection of preserved magnetosome can reach higher PEI, nucleic acid fragments immobilization and ligation efficiency. By the adoption of the solution, cell transfection ratios of Hela, Hep-G2, Cos-7 and C2C12 are respectively higher than cell transfection ratio of magnetosome preserved by a routine method. The BMPs preserving solution provided by the invention is of great practical significance for more efficiently preserving BMPs for longer and more widely performing its application value.

The invention discloses a CHO cellar strain of high-effective expressive exogenous rhBMP2, which is characterized by the following: adopting CHO-dhfr- in the dihydrofolic acid reductase flaw-typed Chinese hamster ovary cell as host cell; infecting exogenous hBMP2 plasmid; adopting secretory expressive product to induce myoblast system C2C12 to display the activity to stimulate sclerotomal cell; obtaining single-clone strain of CHO (rhBMP-2) with the content of BMP2 at 7.83ug / 24híñ106; fitting for clinic treatment.

The invention provides the application of transcription factor MyoD in regulating the expression of pig RTL1 gene. Taking RTL1 as the research object, by constructing the recombinant plasmid of porcine RTL1 gene promoter dual luciferase reporter gene, and finding the core promoter region of RTL1; verifying the interaction between the transcription factor MyoD and the core promoter region of RTL1; then constructing MyoD overexpression vector and synthetic small interfering RNA were used to detect the effect of MyoD on RTL1; the transcription factor MyoD binding site mutant fluorescent expression vector MyoD‑mut was also constructed to detect luciferase activity after transfection into PK15 and C2C12 cells. The invention confirms the influence of transcription factor MyoD on RTL1 gene transcriptional regulation for the first time, brings a deeper level of cognition to the expression regulation of RTL1, and has a good application prospect in the improvement of livestock meat quality traits and the molecular regulation mechanism of muscle development .

The invention provides the application of transcription factor SP1 in regulating the expression of pig RTL1 gene. The present invention takes RTL1 as the research object, by constructing the recombinant plasmid of porcine RTL1 gene promoter dual luciferase reporter gene, and finding the core promoter region of RTL1; and verifying the interaction between the transcription factor SP1 and the RTL1 core promoter region; Then, the SP1 overexpression vector and synthetic small interfering RNA were constructed to detect the effect of SP1 on RTL1; the transcription factor SP1 binding site mutation fluorescent expression vector SP1-mut was also constructed to detect luciferase activity after transfection into PK15 cells and C2C12 cells. The invention confirms the influence of transcription factor SP1 on RTL1 gene transcriptional regulation for the first time, and brings a deeper level of cognition to the expression regulation of RTL1. It is applied to the improvement of livestock meat quality traits and the research on the molecular regulation mechanism of muscle development, and has good application prospect.

The invention relates to applications of dibenzyl citrate in inhibition of calcium ion deposition diseases, and belongs to the technical field of chemical biology. Experimental data of the present invention show that the dibenzyl citrate has effects of good mouse myoblast C2C12 calcification inhibition and effective reduction of calcium ion concentration in human skeletal muscle cell HSK, and after cavia porcellus with arthritis is selected as the animal model and the dibenzyl citrate is used to treat, the bone joint density photo of the cavia porcellus obviously becomes clear.