4199results about "Skeletal/connective tissue cells" 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 method for improving beef quality, an action mechanism and an experimental method, in a cattle body, vitamin A can activate the expression of EBF2 through an active metabolite RA of the vitamin A, and the EBF2 can inhibit the transcription process of CYP26B1 in a targeted manner to maintain the activity of a retinol signal channel, so that PPAR gamma and downstream lipid metabolism related genes thereof are activated, and the activity of the retinol signal channel is improved. Fatty acid transport and lipid accumulation in fat cells in the cattle muscle are promoted, fat deposition in the cattle muscle is promoted, and the beef quality is improved. By constructing the molecular network for regulating and controlling the formation of the fat in the cattle muscle, a complex regulation and control mechanism for controlling the fat deposition in the cattle muscle can be understood more deeply, a method for improving the beef quality based on regulating and controlling the vitamin A mediated key factor EBF2 is provided, and a new way is provided for improving the beef quality in a targeted manner.

The present application relates to chondrocyte compositions and kits comprising morselized cartilage tissue particles and a biodegradable matrix. Applications and methods of using the chondrocyte composition for cartilage grafts, repairing cartilage injuries and defects, and joint repair are provided.

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 invention provides a pourable tumor organoid vascularization model and a construction method thereof, and relates to the technical field of organoid culture. A temperature-sensitive degradable material and a photo-crosslinking modified temperature-sensitive material are used, the composite stent is printed through two channels, a multi-component gel mask partition curing process is combined, the stability of a perfusion pipeline is improved, and a partitioned highly-bionic complex microenvironment is accurately constructed in a macroscopic three-dimensional space; and constructing to obtain the perfusion tumor organ vascularization model. The model is high in bionic degree and stable in structure, can be applied to deepening cell and microenvironment interaction research and tumor-related drug screening, and has a good application prospect.

The invention provides a recombinant humanized elastin with high biological activity as well as a preparation method and application thereof, and belongs to the technical field of genetic engineering and biological materials. 100% of the amino acid sequence of the elastin is derived from human elastin and is prepared by expression of genetically engineered bacteria; the protein has a typical beta-spiral hydrophobic structural domain and a lysine-rich hydrophilic structural domain, has a good bionic structure and functional characteristics, can significantly promote proliferation, adhesion and migration of human skin fibroblasts, also has excellent antioxidant, anti-wrinkle and anti-aging activities, and can be used for preparing a biomimetic protein. The hydrogel can be widely applied to the fields of skin repair dressings, injection implants, artificial skin, biomedical materials, functional skin care products and the like; in addition, the elastin is good in solubility, high in stability, mature in expression system and suitable for large-scale production.

Disclosed are methods of treating traumatic brain injury in a subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

The invention relates to the technical field of tumor biomedical engineering and organ chips, in particular to a vascularized gastric cancer organ chip and a preparation method thereof.The chip is composed of an integrated micro-fluidic main body, an annular micro-column array and an optical sealing film, and a central culture cavity is divided into a tumor area and a blood vessel area by micro-columns; the preparation method comprises the following steps: injecting a fibrous protein solution containing human umbilical vein endothelial cells and cancer-related fibroblasts into a vascular region, and adding thrombin for in-situ gelation to form a pre-vascularized network; the method comprises the following steps: mixing a patient-derived gastric cancer organ with a methacrylic acid esterified gelatin pre-polymerized solution, injecting the mixture into a tumor area, and carrying out photo-crosslinking immobilization; culturing for 7-14 days under the dynamic perfusion condition of a mixed culture medium to obtain a three-dimensional gastric cancer organ model containing a capillary network; the method is expected to be used for rapid, low-cost and high-throughput screening of chemotherapy or anti-angiogenesis drugs and accurate prediction of individual curative effects of patients.

Provided herein are altered cell lines comprising a suspension cell line adapted from an adherent cell line having a different expression profile from a corresponding non-altered adherent cell line, methods for generating altered cell lines, and methods of characterizing altered expression profiles for a gene, a transcript, or a protein in an altered cell line.

Disclosed are methods of treating infertility or increasing fertility of a female subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

Disclosed are methods of treating perianal fistula a subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

The invention relates to a medicine, a medicine composition and application of the medicine and the medicine composition in treatment of liver fibrosis. The medicine and the medicine composition comprise an umbilical cord mesenchymal stem cell membrane; the umbilical cord mesenchymal stem cell membrane is prepared by the following steps: coating the surface of a temperature-sensitive culture dish with a matrix; adding the cell suspension of the umbilical cord mesenchymal stem cells into a temperature-sensitive culture dish for culturing; the temperature is reduced, the umbilical cord mesenchymal stem cells and the extracellular matrix secreted by the umbilical cord mesenchymal stem cells are separated in a lamellar mode, and the umbilical cord mesenchymal stem cell membrane is obtained. The medicine and the medicine composition provided by the invention can effectively relieve liver fibrosis.

The invention relates to methods for culturing cells, and cultured animal cells for use in described method. Also provided is a modified animal cell and guide RNAs targeting genes to be modified in the modified and cultured cells of the invention.

The invention discloses a culture medium and application thereof, and a method for in-vitro separation culture and adipogenic differentiation of precursor adipocytes in goat muscle, and relates to the technical field of cell culture, the culture medium comprises the following components: 8-12 [mu] g / mL of insulin, 8-12% by volume of fetal calf serum, 95-105 U / mL of penicillin, 95-105 [mu] g / mL of streptomycin and the like. On the basis of the prior art, a proper amount of liquiritin and pilocarpine are mainly added into a culture medium, and synergistic cooperation of the liquiritin and pilocarpine is found, so that the adipogenic differentiation capacity of cells such as mesenchymal stem cells and precursor adipocytes is remarkably improved, and the application range is wide. On the basis, the invention also provides a complete set of perfect in-vitro isolated culture and adipogenic differentiation method of the goat intramuscular precursor adipocytes, and a method and a way are provided for a series of researches developed through the goat primary intramuscular precursor adipocytes in the future.

The invention relates to the technical field of biological products, and discloses a mesenchymal stem cell hair growth liquid and a preparation method thereof, and the mesenchymal stem cell hair growth liquid comprises the following components: hair follicle mesenchymal stem cells; a hair follicle mesenchymal stem cell exosome; growth factors: a vascular endothelial growth factor (VEGF), a fibroblast growth factor (FGFs), a keratinocyte growth factor (KGF-7) and interleukin-10 (IL-10); vitamins: vitamin E, vitamin B5, vitamin B2, vitamin B6, vitamin C and biotin; the plant extracts comprise a cacumen biotae extract, a polygonum multiflorum extract, a ginseng extract, a peppermint extract, a purslane extract and a rhodiola rosea extract; other active ingredients: tripeptide and digoxin; the auxiliary material is a hyaluronic acid-collagen peptide graft copolymer; and the balance of a phosphate buffer solution.

The invention belongs to the technical field of biotechnology and animal models, and discloses a double-person-derived mouse model for simulating a tumor immune microenvironment and a construction method and application thereof. The method comprises the following steps: firstly, pretreating NSG immunodeficient mice by adopting low-dose whole-body irradiation in combination with double-antibody targeted bone marrow depletion, and transplanting CD34 + hematopoietic stem cells from the same human donor to complete human immune system reconstruction; separating tumor primary cells, tumor-related fibroblasts and tumor vascular endothelial cells of the same donor, performing three-dimensional co-culture to obtain homologous human tumor organs, and performing in-situ inoculation to immune reconstruction mice to obtain a target model. The core defects of MHC mismatching, low immune reconstruction efficiency, poor tumor immune microenvironment simulation degree, low clinical consistency and the like of an existing model are overcome, and the method can be used for tumor immune treatment drug screening, microenvironment mechanism research and personalized tumor treatment scheme verification.

The invention discloses a double-structural-domain recombinant human fibronectin, the sequence of which is shown as SEQ ID NO.3, and the double-structural-domain recombinant human fibronectin is formed by fusing a collagen binding structural domain and a cell binding structural domain of human fibronectin. The invention also provides a recombinant expression vector containing the protein coding gene and a host cell. The protein is efficiently expressed through eukaryotic cells and engineering bacteria escherichia coli, and a fermentation and purification process suitable for an industrial scale is established in a matched manner. Functional experiments prove that compared with commercially available fibronectin products with a single structural domain, the double-structural domain recombinant human chemical fibronectin disclosed by the invention can remarkably promote proliferation, migration, adhesion and differentiation activity of human skin fibroblasts, and has wide application potential in the fields of skin repair, skincare and biomedical materials.

The invention relates to the technical field of repair materials, and particularly discloses an extracellular matrix fiber material as well as a preparation method and application thereof. The invention discloses a preparation method of an extracellular matrix fiber material. The preparation method comprises the following steps: taking animal tissues, and carrying out virus inactivation, decellularization and antigen removal, degreasing, drying, grinding and sterilization; the decellularization and antigen removal method comprises the following specific steps: sequentially soaking a sample with a surfactant solution for h; then, soaking treatment is carried out by adopting a salt-alkali solution which contains 0.05-3 mol / L of salt and has the pH value of 8-11; then carrying out soaking treatment by adopting a 0.1-1wt% enzyme solution; and in degreasing, one or more of normal hexane, trichloromethane, dichloromethane, methanol, ethanol, acetone, diethyl ether and ethyl acetate are used for soaking the sample. According to the technical scheme, the ECM powder particle material with high growth factor content is prepared.

The invention discloses application of bifidobacterium adolescentis CCFM1302 in promoting synthesis of host collagen, and belongs to the technical field of microorganisms and medicines. The metagen of the bifidobacterium adolescentis CCFM1302 disclosed by the invention has a good effect of regulating and controlling synthesis and degradation of collagen, the moisture and elasticity of the skin are improved, the antioxidant capacity of the skin is improved, the inflammation level of an organism is reduced, and the stable state of collagen is maintained. The method specifically comprises the following steps: in vitro promoting the enzyme activity of HSF cell LH1 by a thallus lysate, inhibiting the enzyme activity of MMP-3, increasing the content of COL I and COL III and the enzyme activity of LH1, P4H and LOX from the outside of a fermented supernatant, and inhibiting the enzyme activity of MMP-3; the inactivated thalli are orally taken to increase the collagen content and the enzyme activity of LH1 and inhibit the enzyme activity of MMP-1 and MMP-3; the fermentation supernatant is orally taken to inhibit enzyme activity of MMP-3 and MMP-1 and degradation of collagen, and the content of TGF-beta is increased.

Disclosed are methods of improving perioral and face fat graft retention in a subject by administering a therapeutic MSC secretome product made by a method comprising culturing bone marrow-derived MSCs under conditions that include oxygen tension below 5% and a culture media with a pH below 7.

The invention relates to the technical field of biological medicines, in particular to rhizoma drynariae extracellular vesicles for treating periodontitis by targeting macrophage ETS2 genes as well as a preparation method and application of the rhizoma drynariae extracellular vesicles. The extracellular vesicles are creatively extracted from the traditional Chinese medicine rhizoma drynariae by combining a differential ultracentrifugation method with a sucrose density gradient centrifugation method, the functions of the extracellular vesicles are studied, and it is found that the extracellular vesicles can effectively slow down alveolar bone resorption in a periodontitis state and reduce periodontal soft tissue inflammation and have a good treatment effect on periodontitis. It is found for the first time that the rhizoma drynariae extracellular vesicles can reduce the inflammatory ability of macrophages in an inflammatory state and can reduce expression of proinflammatory factors TNF-alpha and IL-6. The invention has important scientific research value and clinical significance for further development and application of various traditional Chinese medicine plant extracellular vesicles including rhizoma drynariae extracellular vesicles.

The present disclosure relates to cellular compositions that are modified to introduce a nucleic acid or vector expressing the same. Such compositions may be used to deliver nucleic acid to a target cell and treat disease such as cancer.

The invention relates to the technical field of biological medicines, in particular to an iRGD targeted and anti-fibrosis small molecule dual-modified exosome as well as a construction method and application thereof. The exosome utilizes the specific binding penetration enhancement effect of iRGD (the amino acid sequence is CRGDKGPDC) and a fibroblast high-expression integrin receptor, so that the enrichment efficiency of the exosome at a focus part is improved; the anti-fibrosis micromolecule Let-7a-5p can inhibit a TGF-beta signal channel by blocking Smad2 / 3 phosphorylation, and the anti-fibrosis micromolecule Let-7a-5p and the anti-fibrosis microRNA carried by the exosome generate a synergistic anti-fibrosis effect; the exosome phospholipid bilayer can also protect Let-7a-5p from enzymolysis and prolong the half-life period, and iRGD modification can reduce the non-targeted uptake rate. An animal model verifies that the double-modified exosome has a good skin fibrosis resisting effect, and side effects such as liver and kidney function damage are not found. The invention provides an effective and safe treatment scheme for resisting skin fibrosis.

The present invention provides preparations of MSCs with important therapeutic potential. The MSC cells are non-primary cells with an antigen profile comprising less than about 1.25% CD45+ cells (or less than about 0.75% CD45+), at least about 95% CD105+ cells, and at least about 95% CD166+ cells. Optionally, MSCs of the present preparations are isogenic and can be expanded ex vivo and cryopreserved and thawed, yet maintain a stable and uniform phenotype. Methods are taught here of expanding these MSCs to produce a clinical scale therapeutic preparations and medical uses thereof.

The invention discloses a ready-to-use activated human umbilical cord mesenchymal stem cell as well as a preparation method and application thereof. The preparation method comprises the following steps: carrying out passage on P0-generation cells for three times to obtain P3-generation cells; cryopreserving P3 generation cells, and establishing a cell seed bank; resuscitating the cells in the seed bank for three times of passage to obtain P6-generation cells; and carrying out activation culture on the P6-generation cells, and resuspending the activated P6-generation cells in a cell protective agent to obtain the human umbilical cord mesenchymal stem cell original suspension. The invention also discloses an umbilical cord mesenchymal stem cell cryopreservation preparation. The cryopreservation preparation is obtained by mixing the human umbilical cord mesenchymal stem cell original suspension with a cryopreservation protective agent, sub-packaging and cryopreserving. Experiments prove that the human umbilical cord mesenchymal stem cell cryopreserved preparation prepared by the invention can be stored for a long time, is convenient to store, transport and use clinically, can complete effectiveness and safety index detection within the shelf life, and has the same activity as a fresh preparation.

The invention discloses a bone organ chip loaded with a microelectrode array and a use method, and relates to the technical field of organ chips, the bone organ chip is made of a polydimethylsiloxane material, and the bone organ chip comprises a three-layer structure: a top layer, a middle layer and a bottom layer; the middle layer comprises two blood vessel micro-channels, two osteoblast micro-channels and a neuron micro-channel, the channels are separated through columns and micro-columns, and cell interaction in a bone micro-environment is simulated; a microelectrode array is arranged on the top layer and is used for monitoring electrical activity of neurons in real time; the bottom layer is provided with a pneumatic valve which can control opening and closing of the micro-channel. According to the invention, nerve and blood vessel channels are introduced, a nerve-blood vessel-immunoregulation bone microenvironment is constructed, physiological processes such as osteogenesis-blood vessel formation coupling and the like are embodied, intervals of different sizes can enhance intercellular crosstalk and allow blood vessels and neuron axons to selectively pass through, so that the chip can simulate a complex microenvironment of bone tissues, and the microenvironment of the bone tissues can be simulated; and an efficient and accurate in-vitro model is provided for bone disease mechanism research and drug development.

The present disclosure relates to methods for the production of a cell-based product based on using synthetic promoters or a cellular input-output unit thereof, configured for controlling the expression of at least one nucleic acid sequence of interest, upon sensing dynamic cellular- and / or environmental- state and / or condition, that leads to a desired phenotype of the cells. The present disclosure further provides cell-based products, in particular, animal cell-based meat products (ACBM).

The invention provides a method for producing cell culture meat through 3D scaffold-free co-culture of pseudosciaena crocea stem cell spheres, and belongs to the technical field of preparation of cell culture meat. A method for producing cell culture meat by 3D scaffold-free co-culture of pseudosciaena crocea stem cell spheres comprises the following steps: mixing and inoculating pseudosciaena crocea myoblasts and pseudosciaena crocea precursor adipocytes into a cell culture plate subjected to anti-adhesion treatment for proliferation culture, spontaneously gathering to form cell spheres, and carrying out differentiation culture to obtain the cell culture meat. According to the co-culture mode provided by the invention, high-efficiency myogenic differentiation and adipogenic differentiation of myoblasts and precursor adipocytes of the large yellow croakers can be realized through proliferation culture and differentiation culture, and an in-vivo microenvironment can be effectively simulated, so that cultured meat similar to a traditional meat product in texture and flavor is produced.

The present invention relates to a modified stem cell and an ex vivo method for preparing defined mature cells from genetic programming of said modified stem cell which can be carried out in absence of certain growth factors, as well as a kit allowing to transform stem cells, and the different uses of the mature cells, for example for in vitro drug screening and toxicology or as cultivated meat. The genetic programming involves a molecular switch between two mutually exclusive programs of stem cell expansion and differentiation. The invention may be used for stem cells or other progenitors cells of any type, from any eukaryotic organism, but finds particular application in pluripotent stem cells from humans and livestock animals.