6782results 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 relates to eye drops containing exosomes and a preparation method and application thereof, and belongs to the technical field of eye drop preparation, the eye drops comprise exosome freeze-dried powder, the exosome freeze-dried powder is prepared from the exosomes obtained after mesenchymal stem cells are treated with TNF-alpha and IFN-alpha, and a herba houttuyniae extract, the eye drops provided by the invention can inhibit the phenotype of macrophage M1 and promote the M1 to be converted into the phenotype of M2; human corneal cell proliferation, migration and damage repair are promoted; mouse cornea injury repair is accelerated; the eye drops can be used for treating xerophthalmia, conjunctivitis and keratitis, and can also be used for preventing eye diseases and relieving asthenopia; the eye drops disclosed by the invention contain the exosome obtained after the mesenchymal stem cells are treated by the TNF-alpha and the IFN-alpha, so that the anti-inflammatory ability is stronger; the mesenchymal stem cell exosome and the herba houttuyniae extract are stored in a freeze-dried form, so that the activity is stable, and the storage time is long; in addition, the cordate houttuynia is cooperated with the exosome, so that the effect playing time can be prolonged, and the use frequency is reduced.

To provide a mitochondria function-improving agent or the like which improves a mitochondria function in a skin cell.SOLUTION: A mitochondria function-improving agent that improves a mitochondria function in a cell contains components included in a swallow's nest as an active ingredient. According to each point of view of the present invention, a mitochondria function-improving agent or the like which improves mitochondria function in a skin cell can be provided.SELECTED DRAWING: Figure 3

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 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 invention provides a preparation method of mitochondria-targeted antioxidant hybrid vesicles. The preparation method comprises the following steps: S1, preparing and separating adipose-derived stem cell nano-vesicles; s2, preparing a liposome loaded with dihydromyricetin and distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-triphenylphosphine targeting peptide by an ethanol injection method; and S3, preparing the hybrid nano vesicles loaded with the dihydromyricetin and the distearoyl phosphatidyl ethanolamine-polyethylene glycol 2000-triphenylphosphine targeting peptide. The invention also provides the hybrid vesicles prepared by the method and application of the hybrid vesicles in preparation of refractory diabetes wound treatment drugs. The lipidosome and the adipose-derived stem cell nano-vesicles are hybridized to prepare the hybridized vesicles capable of targeting mitochondria and resisting oxidation, the hybridized vesicles are applied to wound treatment for the first time, the problem of oxidative stress of diabetic wounds is solved, wound healing is accelerated, and a new strategy is provided for optimizing mitochondrial function defects and oxidative stress of the diabetic wounds.

A system for growing cells comprising a bioreactor chamber for growing the cells, a delivery system delivering a perfusion solution to the bioreactor chamber for perfusion of the perfusion solution through the cells, a dialysis system having a dialyzer, a dialysate for performing a dialysis and a filter for reducing ammonia content in said dialysate, and a controller that circulates the perfusion solution through the dialyzer and the dialysate through the filter.

The invention provides a stem cell exosome vesicle induced by traditional Chinese medicine functional components as well as a preparation method and application thereof, and belongs to the technical field of biological medicines. According to the preparation method of the stem cell exosome vesicle, traditional Chinese medicine functional components serve as an exosome inducer, stem cells can be induced to proliferate and secrete exosome with the enhanced function, the yield of the exosome can be increased, bioactive molecules can be loaded, and the bioavailability of the exosome in diabetes treatment is remarkably improved. The stem cell exosome vesicle disclosed by the invention can be applied to treatment of diabetes and complications thereof, not only can improve functions of islet cells, but also can treat chronic inflammation, microangiopathy and other metabolic diseases related to diabetes, and has a wide clinical application prospect.

The invention relates to a dynamic-covalent hybrid cross-linked hydrogel material as well as a preparation method and application thereof. The preparation method comprises the following steps: dissolving a component A-phenylboronic acid modified polymer derivative in a biocompatible medium to obtain a solution A; dissolving a component B, namely a macromolecular derivative containing a vicinal diol group, into a biocompatible medium to obtain a solution B; dissolving a component C-double bond group modified polymer derivative in a biocompatible medium to obtain a solution C; and mixing the solution A, the solution B and the solution C with certain concentrations for a certain time, and performing illumination under a photoinitiator for a certain time to obtain the dynamic-covalent hybrid cross-linked hydrogel material. Compared with single dynamic bond crosslinking or covalent bond crosslinking, the prepared hydrogel material has machinability and mechanical stability; the hydrogel material has plasticity, shear thinning, self-repairing property and the like in the early stage of crosslinking; enough mechanical strength and long-term stability are realized in the later stage of crosslinking.

The invention provides an exosome, a mesenchymal stem cell and application of the exosome and the mesenchymal stem cell in treatment of type 2 diabetes mellitus, and belongs to the technical field of biological medicine. The exosome secreted by the umbilical cord mesenchymal stem cells of the composite pretreatment group is used for treating the type 2 diabetes mellitus, and the obvious advantages are shown. Compared with a model control group, the exosome has the advantages that the glucose tolerance of diabetic rats treated by the exosome is obviously improved, the blood glucose value is reduced by about 47.5% within 120 minutes, the area under a blood glucose curve (AUC) is reduced by about 50.5%, the serum insulin is increased by about 147.7%, insulin secretion is effectively promoted, the glycosylated hemoglobin level is obviously reduced, and the long-term blood glucose control effect is good. By integrating various indexes, the exosome obtained by the composite pretreatment scheme provided by the invention has a remarkable effect of improving the illness state of rats with type 2 diabetes mellitus, provides a new effective strategy for treatment of type 2 diabetes mellitus, and shows a good application prospect.

The invention discloses a porcine fibroblast line capable of being passaged for a long time and application of the porcine fibroblast line. According to the invention, pig fibroblast tissues are taken as a source, pig fibroblasts at different parts are obtained through a tissue block separation method, in-vitro long-term passage is carried out, the cells are triggered to break through the Harike limit, monoclonal screening is further carried out through a limited dilution method, and a pig fibroblast line with a single source is obtained. After screening, porcine fibroblasts with two parts can be subjected to long-term passage in vitro, namely ear edge fibroblasts and rear leg fibroblasts, have collagen secretion capability and are preserved in China Center for Type Culture Collection with preservation numbers of CCTCC NO: C202511 and CCTCC NO: C202512, the cell line breaks through the Harike limit, is already transferred to 42 generations at present, and has the advantages of being capable of secreting collagen, capable of secreting collagen, capable of secreting collagen and capable of secreting collagen. The cells are in a healthy and full long fusiform state, the proliferation level is not influenced after long-term passage, and the method can be applied to preparation of cell culture meat.

The invention provides a serum-free culture medium for in-vitro suspension culture of porcine muscle stem cells and application of the serum-free culture medium. The serum-free culture medium can help the porcine muscle stem cells to be subjected to suspension enlarged culture under serum-free and carrier-free conditions. Compared with a traditional porcine muscle stem cell culture scheme and culture medium, the serum-free suspension culture medium has the advantages that high-density continuous culture of porcine muscle stem cells can be realized in a batch culture process and a fed-batch culture process under the serum-free carrier-free suspension culture condition, high cell viability is maintained, and the survival rate of the porcine muscle stem cells is increased. The peak density of the cells reaches 1 * 10 < 7 > cells / mL or above, and the cell viability is maintained to be 90% or above. The method has the beneficial effects that the problem of serum-free carrier-free efficient suspension amplification of the seed cells of the cell culture meat is effectively solved, and a technical and raw material basis is provided for industrial production of the cell culture meat.

The invention belongs to the technical field of tissue engineering, and particularly discloses a preparation method of a 3D full-thickness skin model. The preparation method provided by the invention comprises the following steps: constructing a corium layer, constructing an inner epidermal layer through first epidermal inoculation, constructing an epidermal layer through second epidermal inoculation and obtaining a complete 3D full-thickness skin model through co-culture. The 3D full-thickness skin model obtained through the preparation method is distinct in gradation, uniform in thickness, free of shrinkage and stable in quality, the epidermal layer and the corium layer are uniformly fused, and the 3D full-thickness skin model is suitable for skin function research and sample evaluation. Meanwhile, the preparation method provided by the invention is simple in process and convenient to operate, does not need additional reagents or anti-shrinkage tools, can meet the requirements of large-scale industrial production, and has good production feasibility and market application prospects.

The invention relates to the technical field of biological medicines, in particular to an umbilical cord mesenchymal stem cell exosome and application of the umbilical cord mesenchymal stem cell exosome in preparation of a medicine for treating male infertility caused by hidden testis. The prepared high-concentration umbilical cord mesenchymal stem cell exosome suspension preparation is directly used for treating cultured spermatogonial cells and spermatoblasts in vitro, is injected into the testis of a mouse, and is used for improving the seminiferous tubule and spermatogenic microenvironment of testis tissue and promoting division and proliferation of the spermatogonial cells and the spermatoblasts. The method can effectively reverse oligospermia and azoospermia caused by the hidden testis, and compared with a model without intervention of treatment means, the method can effectively promote increase and even regeneration of sperms in the testis.

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 application of an enhanced and optimized mesenchymal stem cell exosome in knee osteoarthritis and muscle cartilage injury, and belongs to the technical field of biological medicine. The treatment activity of the umbilical cord mesenchymal stem cell exosome is remarkably improved by combining a composite pretreatment technology with a 3D culture system. Through the synergistic effect of the total flavonoids of rhizoma drynariae and tanshinone IIA, the pathological process of knee osteoarthritis can be regulated and controlled in multiple dimensions, including inflammatory reaction inhibition, cartilage repair promotion and joint function improvement, and the functional limitation of single-component pretreatment is broken through. The invention provides a safe and efficient novel treatment technical strategy for relieving knee osteoarthritis and muscle cartilage injury, and has a wide clinical application prospect.

The invention relates to a medical simulation experiment method and device for acting apoptosis vesicles on primary cartilage cells based on cell line culture, and the method mainly comprises the following steps: S1, co-culture system construction, S2, multi-dimensional observation and analysis, and S3, operation risk assessment. In the step S1, primary cartilage cells, primary immune cells and cell line culture apoptosis vesicles are co-cultured; s2, carrying out multi-dimensional observation and analysis on the influence of apoptosis vesicles on cells; and S3, mainly constructing a multi-dimensional risk scoring system, and carrying out risk grade division by applying a biological network model. The invention aims at simulating and evaluating the influence of cell line culture apoptosis vesicles on primary cartilage cells in vitro and the interaction between the cell line culture apoptosis vesicles and primary immune cells, so that an experimental basis is provided for subsequent clinical application.

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 relates to a cartilage organ constructed on the basis of a DNA-silk fibroin hybrid hydrogel sustained-release system as well as a preparation method and application of the cartilage organ. The DNA-silk fibroin hybrid hydrogel sustained-release system is a hydrogel system formed by printing a DNA-silk fibroin hybrid hydrogel sustained-release system premix solution through a digital light processing system, and the DNA-silk fibroin hybrid hydrogel sustained-release system premix solution contains DNA, silk fibroin, acryloylated RGD peptide, acryloylated polyethylene glycol NHS ester, glucosamine, TD-198946 and 2, 4, 6-trimethyl-1, 3-pentanediol. The invention discloses a lithium 2, 6-trimethylbenzoyl phosphate. Compared with the prior art, the glucosamine and the TD-198946 introduced into the DNA-silk fibroin hybrid hydrogel sustained-release system disclosed by the invention can be used for remarkably improving and promoting the cartilage differentiation efficiency of the bone marrow mesenchymal stem cells; the cartilage graft can be transplanted to a cartilage defect position as a cartilage graft to promote cartilage regeneration and repair.

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.

The invention provides a composition of umbilical cord mesenchymal stem cell exosome combined with GPT hydrogel, a preparation method and application, and belongs to the technical field of wound dressing. The invention solves the problems of fast degradation and low utilization rate in the existing mode of treating the diabetic foot wound surface by purely injecting exosome into the wound surface. The preparation method of the composition of the umbilical cord mesenchymal stem cell exosome and the GPT hydrogel comprises the following steps: S1, culturing to obtain the umbilical cord mesenchymal stem cell exosome; s2, preparing a 4-carboxyphenylboronic acid modified gelatin solution; s3, preparing a tannic acid solution; s4, mixing and stirring the 4-carboxyphenylboronic acid modified gelatin solution prepared in the step 2 and the tannic acid solution prepared in the step 3 to finally obtain the injectable hydrogel containing the exosome. The exosome containing umbilical cord mesenchymal stem cells is combined with the GPT hydrogel microspheres, so that the residence time of the exosome in the wound skin can be prolonged, and a better treatment effect can be achieved.

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 cell differentiation and the technical field of myocardial tissue regenerative medicine, in particular to a method for rapidly inducing directional differentiation of embryonic stem cells into myocardial cells in vitro. A matrix sandwich method is adopted, specific small molecule compounds are added in different time periods to induce the embryonic stem cells to differentiate towards the myocardial cells, and nearly 80% of myocardial cells can be obtained on the seventh day of induced differentiation. Compared with a traditional in-vitro myocardial differentiation induction scheme, the induced differentiation method provided by the invention has the advantages that the variety of related small molecules is few, the dosage is low, the induced differentiation time can be effectively shortened from 15 days to 7 days, the induced differentiation cost is saved, and the method is suitable for large-scale popularization and application. A novel way which is more economical, simpler, more convenient, more stable and more efficient is provided for the production of the stem cell-derived myocardial cells; the cardiac muscle cells obtained by induced differentiation are relatively high in purity and relatively wide in application prospect, and are of great significance to research on drug screening, myocardial regenerative medicine and cardiac developmental biology.

The present invention relates to a method for constructing an eight-gene-edited xenogeneic organ transplantation donor pig, belonging to the field of animal biotechnology. In the wild-type porcine fetal fibroblast cell line, the GGTA1, β4GalNT2, and CMAH genes are knocked out by using the CRISPR / Cas9 gene editing technology, and the humanized genes of hCD39, hCD46, hCD55, hCD59, and hTBM are transfected. Combining with somatic cell cloning technology, GTKO / β4GalNT2KO / CMAHKO / hCD39 / hCD46 / hCD55 / hCD59 / hTBM eight-gene-edited cloned pigs are constructed. Further, through genotype, mRNA, protein expression identification and functional analysis, eight-gene-edited xenogeneic organ transplantation donor pigs are obtained. The present invention solves the technical problems of high production difficulty, low efficiency, and low survival rate of donor pigs for multi-gene-edited xenogeneic organ transplantation, and maximally solves the common problems of immune rejection reaction and complement dysregulation faced during xenogeneic organ transplantation, laying a foundation for more targeted development of donor pigs suitable for different tissue and organ xenotransplantation, and having important value for promoting the clinical transformation of xenogeneic organ transplantation.

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.

The invention provides a cattle FTO gene promoter mutation site and application thereof, and belongs to the field of biotechnology and genetic engineering, the mutation site is located in an FTO gene promoter region on a cattle genome 18 # chromosome, and the mutation site is specifically characterized in that an A base at a transcriptional start site upstream-33 base position is mutated into a G base (chromosome coordinate: Chr: 22040490); according to the mutation, the expression level of the FTO gene is remarkably improved, and the differentiation capacity of intramuscular fat precursor cells is remarkably enhanced; the FTO gene promoter mutation site provided by the invention not only can increase the gene expression quantity, but also can promote the differentiation of intramuscular fat precursor cells, and provides a solid technical basis for the cultivation of new beef cattle varieties and the increase of intramuscular fat content.

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 verification method for regulating and controlling proliferation and differentiation of bovine adipocytes by gene interference, which comprises the following steps: cutting bovine adipocyte tissues into small blocks, directly pasting the small blocks in a culture dish for culture, and promoting cell growth and proliferation by using nutrition and growth factors of the tissues; a ZNF423 gene is targeted by adopting an siRNA technology, siRNA is transfected into bovine fat cells by utilizing a transfection reagent, and the siRNA is an interference fragment containing si-ZNF423; the method comprises the following steps: respectively extracting total RNA (Ribonucleic Acid) and total protein of bovine adipocytes, and detecting by utilizing qPCR (Quantitative Polymerase Chain Reaction) quantitative analysis, a WB (White Batch) technology, EdU dyeing, flow cytometry detection, oil red O, BODIPY (Boron Dipyrromethene), Nile Red dyeing, TG (Triglyceride) content determination and the like. According to the invention, ZNF423 gene expression is interfered by the interference fragment containing si-ZNF423, and the effect of inhibiting proliferation and differentiation of bovine adipocytes is detected by various methods, so that the effect of the interference fragment in the bovine adipocytes is known conveniently, and regulation and control of proliferation and differentiation of the bovine adipocytes are realized by interfering gene expression by the interference fragment. The beef quality can be improved; the beef cattle breeding can be optimized.

The present disclosure relates to methods, cell culture medium and compositions that promote cell proliferation during fetal bovine serum free cell culture.