3232results about "Skeletal/connective tissue cells" patented technology

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 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.

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 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 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 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 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.

The invention relates to injectable temperature-sensitive hydrogel loaded with traditional Chinese medicine active exosomes, and belongs to the technical field of clinical treatment of diabetic foot refractory wounds, and the injectable temperature-sensitive hydrogel is obtained by poloxamer 407 hydrogel and a hyaluronic acid loaded composite carrier; the composite carrier body is formed by coating a traditional Chinese medicine active exosome with a coating solution; the traditional Chinese medicine active exosome is obtained by inducing adipose-derived mesenchymal stem cells with a modified astragalus extract. The injectable temperature-sensitive hydrogel is obtained by loading a composite carrier with poloxamer 407 hydrogel and hyaluronic acid, the poloxamer 407 and the hyaluronic acid quickly form a physical barrier on a wound surface after being injected, a moist healing environment is provided, and continuous and slow release of exosome and active ingredients is realized by means of the temperature-sensitive characteristic and the network structure of the poloxamer 407 and the hyaluronic acid, so that the wound healing effect is improved. And over-high local concentration or premature loss caused by sudden release of the medicine is avoided.

The invention discloses a method for quickly and efficiently differentiating iPSC (induced pluripotent stem cells) into mesenchymal stem cells in a serum-free manner, the method comprises the following steps: firstly inducing the iPSC into a mesoderm precursor, then transferring the mesoderm precursor into a liquid culture medium for suspension culture, and finally obtaining iMSC (iPSC-doped MSC) by using an enrichment culture medium, and the method can shorten the differentiation time, improve the purity of the iMSC and obtain a large number of homogenized iMSCs.

The invention discloses an adipose-derived stem cell exosome compound loaded with silver nanoparticles and application of the adipose-derived stem cell exosome compound loaded with the silver nanoparticles, and ADSC-Exos loaded with the silver nanoparticles is developed by integrating AgNPs into an exosome of ADSCs. In vivo, the Exo-AgNPs can accelerate healing of infected wounds by reducing bacterial colonization and promoting collagen deposition, angiogenesis and M2 type macrophage polarization. In vitro, the Exo-AgNPs activates the functions of fibroblasts and vascular endothelial cells, and shows an excellent antibacterial property. In addition, the Exo-AgNPs induces polarization of M2 type macrophages and inhibits secretion of proinflammatory cytokines by activating autophagy. The results show that Exo-AgNPs is expected to become an ideal biological material for treating infected wounds.

The invention belongs to the technical field of exosome freeze-dried powder, and discloses exosome freeze-dried powder and a preparation method thereof, the exosome freeze-dried powder contains mesenchymal stem cell exosome, trehalose, sodium hyaluronate, hydroxypropyl-beta-cyclodextrin, glutathione, ceramide and fructo-oligosaccharide. According to the exosome freeze-dried powder, hydroxypropyl-beta-cyclodextrin, glutathione, ceramide and fructo-oligosaccharide are added, the membrane structure integrity of the exosome is effectively protected, the retention rate of active substances in the exosome freeze-dried powder is high, and therefore the effects of resisting inflammation, regulating cell metabolism, promoting tissue repair and the like of the exosome can be fully exerted. In addition, glutathione can directly remove skin free radicals and inhibit oxidative stress injury, and has an anti-oxidation effect; ceramide can also supplement skin lipid barrier, enhance the water locking capacity and enhance the moisturizing effect of sodium hyaluronate; fructo-oligosaccharide can also regulate skin micro-ecological balance, and has anti-inflammatory and bacteriostatic effects.

Provided are chemically modified silk fibroin and a use thereof in cell and organoid culture. The chemically modified silk fibroin is a product obtained by sequentially carboxylating silk fibroin and modifying same by an organic amine having a phenol group. Also provided is a hydrogel based on the chemically modified silk fibroin. A hydrogel system has a hierarchical structure and mechanical properties similar to those of an extracellular matrix, and exhibits characteristics such as definite composition, controllable physicochemical properties, low cytotoxicity, good biocompatibility, and biodegradability, can support the growth and differentiation of cells and organoids, and is suitable as a matrigel for cell and organoid culture.

The invention discloses an MSC culture medium composition and a culture method, and belongs to the field of biological medicine. Aiming at the problems of low MSC amplification efficiency and poor function maintenance in the prior art, the invention provides the MSC culture medium combination, which comprises a basic culture medium and functional factors, and the functional factors comprise an activation functional factor, an amplification functional factor and a steady-state functional factor which are sequentially added according to a preset time interval; wherein the activating functional factors comprise b-FGF, EGF and TGF-beta 1; the amplification functional factors comprise b-FGF (Fibroblast Growth Factor), PDGF-BB (Platelet Derived Growth Factor The steady-state functional factors comprise a b-FGF (Fibroblast Growth Factor) and IGF-1. The cell cycle is quickly started by activating the functional factors, differentiation is inhibited, and the functional factors are amplified to promote large-scale cell amplification; steady-state functional factor equilibrium amplification and dryness maintenance are carried out; and different functional factors are sequentially added according to a preset time interval, and the concentration of the factors is dynamically adjusted, so that differentiation tendency or aging acceleration caused by long-time high-concentration stimulation of a single factor is avoided, long-term efficient amplification of the MSC is realized, and the multifunctionality of the MSC is kept.

Provided herein are methods and related compositions for enhancing or enhanced partial reprogramming of target cells in a subject in need thereof (e.g., a human subject suffering from or at risk of a disease), the method comprising administering a plurality of mesenchymal lineage progenitor or stem cells (MLPSCs), exosomes derived therefrom, or conditioned culture media derived therefrom to a subject that expresses or will express one or more reprogramming factors in a population of target cells, whereby a plurality of the target cells in the subject become partially reprogrammed, but not fully reprogrammed.

The invention relates to the technical field of rhinitis spray preparation, in particular to a method for preparing allergic rhinitis spray from umbilical cord mesenchymal stem cell exosomes, which comprises the following steps: 1, umbilical cord treatment; 2, culturing umbilical cord mesenchymal stem cells; 3, preparing an umbilical cord mesenchymal stem cell exosome; and 4, preparing the rhinitis spray. The rhinitis spray prepared by the method disclosed by the invention is simpler and more convenient to use. The exosome, the platelet growth factor and other substances in the rhinitis spray well solve the core problems of single curative effect, large side effect, low compliance and the like in the traditional rhinitis treatment, and through the induction and repair functions of the stem cell exosome and the bactericidal ability of the antibacterial peptide, the nasal cavity environment is improved, the anaphylactic reaction is reduced, the nasal mucosa is repaired, and the rhinitis treatment effect is improved. The allergic rhinitis is expected to be fundamentally treated.

The invention discloses a sleep-promoting nasal spray based on mesenchymal stem cell-derived exosomes and preparation and application thereof.The preparation method comprises the steps that 1, mesenchymal stem cells separated from umbilical cord tissue are cultured to P3-P5 generations, cell culture supernate is collected, cells are digested, and then cell debris and macromolecular impure protein are removed through pretreatment; (2) filtering and concentrating the pretreated cell culture supernatant and replacing a part of solution; (3) carrying out degerming filtration on the solution obtained in the step (2) through a 0.22 [mu] m filter membrane to obtain an MSC-Exos solution; and (4) adding a stabilizer with a proper concentration into the MSC-Exos solution to obtain a compound MSC-Exos solution, namely the sleep-promoting nasal spray. The product is a cell-derived natural active substance, can significantly improve sleep, and does not involve any pharmaceutical ingredient.

The invention discloses a culture method of high-activity human urine-derived stem cells, which relates to the technical field of biological medicine, and is technically characterized by comprising the following steps: collecting urine samples of pregnant and lying-in women, coating a culture plate with gelatin, inducing adherence, performing passage amplification, performing immunofluorescence identification, performing function detection and other optimized conditions, and separating and culturing the stem cells. Compared with urine-derived stem cells of normal adults, the stem cells derived from pregnant women have stronger activity, higher dryness and faster multiplication capacity. The invention provides a novel and superior cell source for tissue engineering and regenerative medicine.

The invention provides an extraction method of umbilical cord mesenchymal stem cell exosomes. The extraction method comprises the following steps: S1, collecting umbilical cord mesenchymal stem cell culture supernatant; s2, centrifuging the culture supernatant under a centrifugal force of 250-350 g, and collecting the supernatant to obtain a first mixed solution; s3, centrifuging the first mixed solution under a centrifugal force of 1500-2500 g, and collecting a supernatant to obtain a second mixed solution; s4, centrifuging the second mixed solution under a centrifugal force of 9000-11000 g, and collecting a supernatant to obtain a third mixed solution; and S5, filtering the third mixed solution, centrifuging under the centrifugal force of 90000-110000 g, collecting the precipitate, adding a PBS buffer solution, centrifuging under the centrifugal force of 90000-110000 g, and collecting the precipitate to obtain the umbilical cord mesenchymal stem cell exosome. The umbilical cord mesenchymal stem cell exosome with the purity up to 99% is extracted from the culture supernatant through twice low-speed centrifugation and three times of high-speed centrifugation.

Provided are immortalized fish fibroblast cells and food products comprising said fish fibroblast cells.

Disclosed is a composition for treating dystrophic epidermolysis bullosa, the composition containing, among other things, a composition which includes cells that are derived from the lesions of dystrophic epidermolysis bullosa patients and that have been genetically engineered such as to produce type VII collagen.

The invention relates to the technical field of biology, in particular to application of active polypeptide in maintaining dryness of umbilical cord mesenchymal stem cells. The active polypeptide disclosed by the invention is PNC-27, and the amino acid sequence of the PNC-27 is as shown in SEQ ID NO: 1. Experiments find that the PCN-27 polypeptide can effectively promote proliferation of the umbilical cord mesenchymal stem cells by regulating expression of periodic proteins Cyclin-D1 and CDK1, and can also effectively maintain dryness of the umbilical cord mesenchymal stem cells by promoting expression of dry gene Oct-4 protein in the umbilical cord mesenchymal stem cells. Experiments fully prove that the PCN-27 polypeptide can effectively maintain the dryness of the umbilical cord mesenchymal stem cells and stimulate the potential application value of the umbilical cord mesenchymal stem cells in the fields of cell culture, biomedicine and the like.

The invention relates to a serum-free culture medium for culturing mesenchymal stem cells as well as a preparation method and application of the serum-free culture medium. Wherein the serum-free culture medium comprises a complete culture medium and praeruptorin E, and the content of the praeruptorin E in the serum-free culture medium is 0.25-3 [mu] M. The inventor finds that a serum-free culture medium containing a certain concentration of praeruptorin E can effectively improve the decline of replicative activity of mesenchymal stem cells in in-vitro passage. Specifically, the serum-free culture medium containing the praeruptorin E with a certain concentration can reduce the expression level of a marker beta-galactosidase with reduced replicative activity in the mesenchymal stem cells with reduced replicative activity after multiple in-vitro passage, reduce the expression of a gene CDKN1A with reduced replicative activity, and increase the multiplication capacity of the mesenchymal stem cells; the mitochondrial damage is reduced.

The invention discloses a chicken myoblast and chicken fat precursor cell co-culture synchronous differentiation method and application thereof, and belongs to the technical field of animal cell co-culture and differentiation. The method comprises the following steps: respectively carrying out in-vitro multiplication culture on chicken myoblasts and chicken fat precursor cells, carrying out mixed co-culture, inducing for at most two days by adopting horse serum with the volume percent of 0.5-2%, carrying out pre-differentiation, and continuously carrying out induced differentiation culture for at most one day by using an induced adipogenesis culture medium. According to the synchronous differentiation method for co-culture of the chicken myoblasts and the chicken fat precursor cells, a cell co-culture system is constructed, time sequence induced differentiation is achieved, and a final product with fat droplets distributed in the gaps of muscle bundles (the average distance is 20-50 microns) and capable of recarving natural chicken intermuscular fat textures is obtained. According to the method, the differentiation period is shortened to be within 3 days from 21 days (the efficiency is improved by 600%); the induction cost is reduced by 42.7%; the final product breaks through the sensory acceptance barrier through bionic structure recarving.

The invention discloses a gelatin microcarrier lysate as well as a preparation method and application thereof. The gelatin microcarrier lysis buffer comprises lyase, a cell protective agent, a cell energy metabolism protective agent, an antioxidant, a buffer solution and a chelating agent. The gelatin microcarrier lysate has extremely strong protectiveness to cells, abandons damaging trypsin and EDTA, and innovatively adds a cell protection combination of a cell protective agent L-arginine, a cell energy metabolism protective agent sodium pyruvate and an antioxidant vitamin C, so that the cell survival rate (gt; 98%) and the retention rate (gt; the method is high in cell yield, good in safety and wide in range.