5809results about "Culture process" 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 a stem cell bioactive nano-vesicle with efficient inflammation regulation and tissue repair effects, the average particle size of the nano-vesicle is 100-350nm, the polydispersity index (PDI) is 0.05-0.40, and the expression rate of phosphatidylserine (PS) on the surface of the nano-vesicle is greater than 40%. According to the preparation method, stem cells are subjected to apoptosis induction, and then a programmed serial extrusion process is adopted to prepare the nano-vesicles. The nano vesicles are moderate in particle size, uniform in distribution and high in production efficiency, the preparation process is easy to operate, the quality is stable and controllable, and the nano vesicles have excellent technical effects in the aspects of repair and regeneration of inflammation injury type tissues and organs.

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 relates to a preparation method of a tricholoma matsutake fermentation product with anti-wrinkle and firming effects as well as a product and application of the tricholoma matsutake fermentation product. The preparation method comprises the following steps: mixing a composite plant extract, a carbon source, inorganic salt and water, then sterilizing to obtain a sterilized culture medium, inoculating a composite probiotic agent into the medium, and carrying out fermentation culture to obtain the tricholoma matsutake fermentation product with anti-wrinkle and firming effects. Centrifuging, filtering and sterilizing the culture product to obtain a tricholoma matsutake fermentation product; the bacterial strains in the compound probiotic agent comprise a combination of streptococcus thermophilus, a lactobacillus paracasei YK-L003 bacterial strain and a saccharomyces cerevisiae YK-S001 bacterial strain. According to the invention, specific strains are subjected to compounding and combined fermentation, and the fermentation product filtrate obtained after fermentation treatment of the three strains has an excellent skin conditioning effect, can effectively improve the skin state and resist and weaken wrinkles, and has an excellent effect when being applied to moisturizing and anti-wrinkle cosmetics.

The invention discloses a serum-free culture system and culture method of NK cells. The invention firstly discloses a serum-free culture system of the NK cells, wherein the serum-free culture system comprises a coating solution, an amplification culture medium, an activation culture medium and autologous plasma; the invention further discloses a culture method for NK cells by using the serum-free culture system. The method for culturing the NK cells by using the serum-free culture system disclosed by the invention reduces the culture cost of the NK cells, can simply, quickly, efficiently and safely obtain a large number of NK cells with high activity, high purity and strong killing ability in vitro, provides a better cell source for clinical application of the NK cells, and promotes further development of an NK cell treatment technology.

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 organoid culture, and particularly relates to a co-differentiated vascularized liver organoid and a construction method thereof. Aiming at the problems that the existing liver organ vascularization mostly adopts a co-culture mode, the operation process is complicated, the cell source heterogeneity is relatively high, the randomness is strong, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized liver organ. The preparation method comprises the following steps: sequentially inducing and culturing directional endoderm cells, posterior preintestinal cells and hepatic endoderm cells, adding an S1 culture medium containing VEGF to induce and culture for 1-3 days, then adding an S2 culture medium containing VEGF, BMP4 and FGF2 to induce and culture for 3-5 days, and then adding an S3 culture medium containing VEGF to induce and culture for 14-20 days to obtain the co-differentiated vascularized liver organ. According to the method disclosed by the invention, the operation process is simplified, the experiment efficiency is improved, the liver organ which is rich in blood vessel network, high in maturity and of a blood vessel-bile duct structure is obtained, and a foundation is laid for large-scale production and clinical application.

The invention belongs to the technical field of stem cells, and particularly relates to a product containing mesenchymal stem cells, a preparation method, a quality control method and application in prevention and treatment of digestive system diseases. According to the preparation method, 1-3 complete culture media are provided for culturing the mesenchymal stem cells, and the obtained mesenchymal stem cells are good in subpackage form; cD73, CD90 and CD105 are highly expressed, and CD19, CD34, CD45, CD11b and HLA-DR are not expressed; the cell proliferation capacity is strong, the osteogenesis and adipogenic differentiation capacity is strong, and the medicine is safe and free of toxic and side effects. The prepared mesenchymal stem cells are high in quality, have good performance of secreting HGF, KGF and IGF-1, and can effectively treat digestive system diseases.

The invention provides a cognitive function in-vitro simulation method based on a microelectrode array (MEA) and brain organs and application of the cognitive function in-vitro simulation method, and belongs to the crossing field of brain organ in-vitro model construction, neural engineering, brain-like intelligence and the like. In an in-vitro environment, external signal (such as sound) information is converted into electrical stimulation input capable of being perceived by brain organs in an MEA electrical stimulation mode, so that the brain organs gradually learn and identify external signals in repeated electrical stimulation; after elution stimulation for a period of time, the reduction condition of recognition accuracy is observed, namely, the important memory and forgetting process in the cognitive function is simulated. According to the method, the electrophysiological response and the biological mechanism change of the brain organs under external electrical stimulation can be quantitatively evaluated, and a new possibility is provided for exploring the advanced cognitive function by utilizing the brain organs.

A recombinant humanized collagen type I alpha-1 (rhCol1A1), and an expression vector and use thereof are provided, belonging to the technical field of bioengineering. The rhCol1A1 has an amino acid sequence of SEQ ID NO: 1. This aim is to solve the problem of difficulty in expressing soluble full-length humanized collagen in Escherichia coli prokaryotic expression systems, and differences between the recombinant collagen and humanized collagen in yeast expression systems. It is proposed that the rhCol1A1 chain is expressed in Expi293F human embryonic kidney cells, and the protein expression level is increased using a fusion expression plasmid, enabling the expression of full-length humanized collagen type I alpha 1 chain. Activity studies have shown that the expressed rhCol1A1 exhibits a desirable activity in promoting cell migration, and may be widely used in the fields of biomedicine, tissue engineering, skin care, and food.

The invention discloses a preparation method and application of a cell culture liver based on a microcarrier. The method comprises the steps of obtaining liver seed cells, preparing the microcarrier, culturing cells in vitro based on the microcarrier, preparing the cell culture liver and the like. The preparation method of the cell culture liver based on the microcarrier is provided for the first time, and the preparation scheme that a cell-microcarrier compound serves as a structural unit and is assembled into the cell culture liver after being mixed with an edible material is adopted, so that on one hand, simulation of a real liver lobule structure can be achieved, and on the other hand, simulation of the liver lobule structure can be achieved; the whole structure relation of liver lobules and connective tissues in liver tissues can be simulated, full reduction of animal liver appearance and structural characteristics is achieved, edible materials used in the preparation process are low in price and good in biocompatibility, and the preparation method is suitable for large-scale industrial production. Liver seed cells can be subjected to efficient amplification and albumin synthesis through three-dimensional culture on a microcarrier prepared on the basis of edible materials, so that the cell culture liver is close to a real animal liver in the aspect of nutrition.

The invention discloses a 3D intestinal organ differentiation method based on human pluripotent stem cells and an induction culture medium and application thereof, and relates to the technical field of stem cells. According to a culture medium formula combination, intestinal organs can be differentiated into various cell types such as epithelial cells, neuroendocrine cells and endothelial cells; the method is a key mark for successful differentiation and functional maturation of intestinal organs. According to the 3D intestinal organ differentiation method disclosed by the invention, histological structures such as intestinal crypts are differentiated from intestinal organs generated by differentiation, and the intestinal organs can creep in a maintenance stage, so that the intestinal organs are changed from structural bionics to functional simulation, and the significance of the 3D intestinal organ differentiation method is far better than that of pure morphological simulation. Through a systematic culture medium formula, a clear operation process and a multi-stage induction strategy, the 3D intestinal organ with structural integrity, cell diversity and functional activity is successfully constructed, and the system provides an efficient, reliable and extensible in-vitro model platform for intestinal biological research and related application.

The invention relates to organoid culture. The invention provides a human epidermal organoid differential medium, a method for constructing an epidermal model and application of the human epidermal organoid differential medium, the human epidermal organoid differential medium comprises a basic medium component and an additive, and the additive comprises lipid. According to the method, the organoid is directly used for constructing the epidermal model, so that the problem of few cell sources is solved; a lipid component is added into the epidermal organ differentiation culture medium, so that the epidermal organ can enhance the barrier function of the cuticle, further differentiation of the epidermal organ can be effectively promoted by utilizing the epidermal organ differentiation culture medium, and a human epidermal model with a basal layer, a spinous layer, a granular layer and a cuticle which are completely morphologically differentiated is obtained. And the operation is simple, large-batch production is easy to realize, and a reliable tool is provided for in-vitro observation, interference of skin disease generation, injury repair and other life activities.

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.

Provided are compositions and methods that enable faithful recapitulation of hair follicle stem cells (HFSCs) identity in culture using a cocktail of factors that include use of an inhibitor of protein kinase C (PKCi), and all-trans retinoic acid (atRA), that successfully target lineage plasticity.

The invention relates to a culture method for activating NKT cells by DC cells in vitro and application of the NKT cells. The preparation method of the NKT cell comprises the following steps: activating a DC cell by using cell inducing factors such as alpha-GalCer, GM-CSF and IL-4, co-culturing the DC cell and a cryopreserved and resuscitated suspension cell left after adherent sorting, and activating and amplifying the NKT cell. The proportion of NKT (CD3 + CD56 +) prepared by the invention is more than 80%, and a small amount of iNKT cells are contained at the same time.

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 belongs to the technical field of biological medicines, and particularly relates to a sperm injury repair model and an establishment method thereof. Comprising the following steps: step 1, establishing a sperm lead poisoning model; and 2, establishing a lead poisoning sperm repair model. The method for establishing the lead poisoning sperm repair model in the step 2 comprises the following steps: (1) preparing a preparation for optimizing sperm functions in an extracellular vesicle way; (2) repairing the damaged sperms by using the preparation for optimizing the sperm function through the extracellular vesicles; and (3) detecting the activity of the repaired sperms. In the invention, by establishing a sperm injury repair model and adopting an extracellular vesicle modification technology, the key functional protein is introduced into the extracellular vesicle, and the method has important significance on auxiliary researches such as development and research of related drugs for improving sperm injury and improving sperm motility of asthenospermia patients in vitro.

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

The invention discloses a method for inducing umbilical cord mesenchymal stem cells to differentiate insulin-like cells and application of the method, purified umbilical cord mesenchymal stem cells are reasonably cultured by adopting culture media with different substances, and the umbilical cord mesenchymal stem cells can be efficiently and stably induced to be differentiated into insulin-like cells. The percentage of insulin positive cells can reach 80%, and the cell preparation prepared after the umbilical cord mesenchymal stem cells are induced can be used for treating clinical diabetics and is high in curative effect.

The invention relates to a cochlea spiral neuron in-vitro organ as well as a construction method and application thereof, and provides a method for obtaining a mature inner ear neuron by utilizing human pluripotent stem cell differentiation in vitro and further constructing an inner ear neuron in-vitro organ on the basis of the mature inner ear neuron. The internal ear neuron in-vitro organoid function stage obtained by the method and the application thereof are also provided.

A brain organoid containing optic vesicles generated based on H9 induction and an eye-brain fusion culture method thereof are provided. H9 embryonic stem cell induction is used to generate a brain organoid containing optic vesicles with primitive visual field. Based on an established optic vesicle brain organoid culture system, microscopic imaging is combined with specific marker antibodies related to early retinal development and photoreceptor cell maturation to structurally and functionally identify the brain organoid containing optic vesicles.

Disclosed are methods, means, and compositions of matter useful for the stimulation of angiogenesis directly by administration of membrane vesicles, such as fibroblast-derived exosomes, and / or through induction of angiogenic cytokines from blood cells contacted with fibroblast-derived exosomes. The invention provides means of treating conditions in which angiogenesis is beneficial through local or systemic administration of exosomes, including those derived from fibroblasts, wherein the fibroblasts are cultured under basal conditions or conditions of hypoxia. In other embodiments exosomes derived from fibroblasts are utilized to augment endogenous regenerative processes, such as hematopoiesis, angiogenesis and neurogenesis, as well as augment regenerative processes stimulated by administration of exogenous therapeutics such as cells, growth factors, or genes.

The invention discloses a method for improving lipid droplet generation and promoting the differentiation process in the bovine adipocyte differentiation process, which comprises the following steps: (1) recovering primary bovine adipocytes, and culturing in a complete growth culture medium; (2) performing induced differentiation on the bovine adipocytes in an induced differentiation culture medium; and (3) replacing the bovine adipocytes subjected to induced differentiation with a culture medium containing 50-100 [mu] mol / L of linoleic acid, and culturing the bovine adipocytes. It is found for the first time that after bovine adipocyte differentiation is finished, culture continues to be conducted in a culture medium added with LA, lipid droplet generation in the bovine adipocyte differentiation process is improved, and the differentiation process is promoted.

The invention provides a gamma delta T cell preparation as well as a preparation method and application thereof, and belongs to the technical field of T cells. The liposome is prepared from the following raw materials in parts by weight: 15-20 parts of modified gamma delta T cell liposome, 1-3 parts of cell stimulating factors and 4-7 parts of culture medium freeze-dried powder, the modified gamma delta T cell lipidosome is prepared by embedding gamma delta T cells through lipidosome, coupling with acetylenic bond modified chitosan, and further mixing with MFAP4 protein and b4GALT1 protein. The culture medium freeze-dried powder is prepared by freeze-drying the culture medium in the engineering culture process of gamma delta T cells, and the cell stimulating factors are resveratrol and quercitrin. According to the gamma delta T cell preparation prepared by the invention, the survival ability and resistance of gamma delta T cells are improved, the anti-tumor activity of the gamma delta T cells is improved, and the gamma delta T cell preparation has relatively good antioxidant and anti-inflammatory effects, reduces the inhibition of inflammation on an immune system and has a very good treatment effect on pancreatic ductal adenocarcinoma.

The invention discloses a filling and repairing agent for promoting angiogenesis and a preparation method thereof, the filling and repairing agent is composed of a vascular endothelial cell supernatant, acellular matrix particles, a suspending aid and a protective agent, the vascular endothelial cell supernatant is prepared by inducing vascular endothelial cells into a tube under a low-oxygen three-dimensional condition, collecting a cell culture solution, centrifuging and concentrating; the acellular matrix particles are obtained by acellular tissue matrix, shearing, freeze-drying, liquid nitrogen grinding and collecting; according to the filling repairing agent for promoting angiogenesis, the vascular endothelial cell supernate and the acellular matrix particles are combined, the filling repairing agent contains high-concentration angiogenesis-related factors and a stable three-dimensional bionic scaffold structure, function complementation is achieved through the synergistic effect of the bioactive factors and the bionic scaffold, and the filling repairing agent for promoting angiogenesis is obtained. Therefore, the filling repair agent which can promote angiogenesis, construct a physical support framework and promote tissue repair and regeneration capacity is innovatively obtained.

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 discloses a cord blood-derived regulatory T cell amplification culture medium and a use method thereof, and relates to the field of animal cell culture and immune cell therapy, the culture medium is a serum-free culture medium, the culture medium is composed of a basic culture medium, a serum substitute combination, a combination of four cell factors, an immunosuppressor, an apparent stabilizer and an antioxidant, and all the components play a role synergistically; according to the use method, cell amplification is realized through a collaborative process of staged environmental regulation and targeted activation. According to the method, the problems of exogenous pollution and batch difference caused by dependence of a serum-containing system on cord blood-derived regulatory T cell amplification in the prior art are solved, the defect that a serum-free scheme is difficult to consider the amplification efficiency and the function stability at the same time is overcome, the cell amplification quality and the clinical application safety are guaranteed, and the method is suitable for large-scale clinical transformation requirements.