8271results 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 an adipose-derived stem cell composition for wound repair as well as a preparation method and application thereof, and belongs to the technical field of stem cells. The method comprises the following steps: adding fetal calf serum, dexamethasone, penicillin, streptomycin, collagen, magnetic particles and a P substance into a DMEM culture medium to prepare a culture medium, performing low-frequency magnetic field culture and separation on adipose-derived stem cells to obtain a stem cell solution, performing embedding to obtain stem cell microspheres, freeze-drying supernate to obtain a paracrine culture substance, and performing freeze-drying on the paracrine culture substance to obtain the adipose-derived stem cells. The adipose-derived stem cell composition for wound repair is prepared by mixing the adipose-derived stem cell with stem cell microspheres, hydroxypropyl methyl cellulose, carboxymethyl chitosan, hyaluronic acid, glycerol and stem cell microspheres, has better effects of implanting into damaged tissues, releasing nutrient substances, promoting neovascularization, regulating oxidative stress and inducing anti-inflammatory response, is beneficial to wound repair, and has a good application prospect. The gel can be converted into a gel phase at the skin epidermis temperature of 35-37 DEG C, components are stabilized, and sustained and controlled release is realized.

The invention provides a large-scale culture method of NK cells and an adopted culture medium, and belongs to the technical field of NK cell culture. And adding fetal calf serum, penicillin, streptomycin and the stimulation amplification promotion magnetic microspheres into the ACTM culture medium, and uniformly stirring and mixing to obtain the culture medium. The large-scale culture method of the NK cells can efficiently activate and amplify the NK cells, is free of exogenous serum and autoserum, safe and effective, can well promote proliferation of the NK cells and prolong the activity of the NK cells, is safe and free of side effects, and has a wide application prospect.

There is provided herein methods generating a population of primed mesenchymal stem cell-derived exosomes. A method in accordance with the disclosure may comprise expanding a population of mesenchymal stem cells (MSCs) in culture, administering one or more priming agents in the culture to prime the population of MSCs and obtain a population of primed MSCs: growing the population of die primed MCSs in culture to produce a primed-MSC-derived conditioned medium; collecting the primed MSC-conditioned medium; and purifying a population of exosomes from the primed MSC-conditioned medium. The one or more priming p agents may comprise a conditioned media derived from a population of stem cells different from the population of MSCs, a Nrf2 activator, or a combination thereof. There is also provided herein methods of treating tissues such as cornea and liver using the population of exosomes from the primed MSC-conditioned medium.

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 present disclosure provides methods of differentiating pluripotent stem cells, including induced pluripotent stem cells, into lineage-specific floor plate midbrain progenitor cells, determined dopaminergic neuronal progenitor cells, committed dopaminergic neuronal progenitor cells and / or dopaminergic neuronal cells. Also provided are compositions uses thereof, such as for treating neurodegenerative diseases and conditions, including Parkinson's disease, and articles of manufacture and kits for use thereof.

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 a culture medium for immune cells as well as a preparation method and application thereof. On the basis of a serum-free culture medium, a specific additive is added into the culture medium and is used as a nutrient substance for growth of immune cells, so that the concentrations of various components in the final culture medium are fixed, the culture efficiency of the immune cells is improved, the cell activity is increased, and the repeatability of subsequent experiments is ensured. Meanwhile, due to the use of a specific additive, animal serum protein added into a conventional culture medium is avoided, so that the risk of infection of exogenous viruses and pathogenic factors is avoided, the immune cells obtained by culture have no sensitization, and the condition that a receptor is allergic to exogenous serum is avoided. In addition, the cost of the immune cell culture medium is reduced, and the preparation process of the culture medium is simplified. Compared with the prior art, the additive provided by the invention can be preserved for a long time in a low-temperature environment, the activity is kept unchanged, and the additive can be taken and used when a culture medium is prepared, so that the preservation cost of immune cells is effectively reduced.

The purpose of the present invention is to provide a culture medium in which cells to be used for producing a cultured meat can be grown, by adding thereto a food material as an additive. As a result of culturing cells using a food ingredient as an additive, it became possible to obtain high growth potential in fibroblasts and myoblasts by adding egg white or a dried product of egg white as a cell growth promoter. Accordingly, the present invention pertaining to a cell-growing culture medium containing a basic culture medium and egg white as a cell growth promoter was accomplished.

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 relates to the technical field of biological tissue engineering, and discloses a nerve-blood brain barrier co-culture model, which comprises a culture chip, and nerves and blood brain barriers respectively constructed in upper culture micropores and lower culture micropores of the culture chip. Wherein a semi-permeable membrane is arranged between the lower-layer culture micropores and the upper-layer culture micropores of the culture chip; the nerves are obtained by inoculating neurons / brain organs obtained by stem cell differentiation into the upper culture micropores for co-culture, and the blood brain barrier is obtained by inoculating brain microvascular endothelial cells obtained by stem cell differentiation into one side of the semipermeable membrane in the lower culture micropores for co-culture. The co-culture model is higher in biomimetic property, and can better simulate the structure and function of the blood brain barrier, for example, the co-culture model has a barrier function closer to the physiological level, the selective permeability of a small molecule drug is matched with the in-vivo permeation condition, and the co-culture model has the function of a small molecule related transporter. The invention further discloses a construction method and application.

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 provides a high-activity product obtained based on cornu cervi pantotrichum stem cells as well as a preparation method and application thereof. The preparation method comprises the following steps: preparing a serum-free culture medium, unfreezing immortalized cornu cervi pantotrichum stem cells, centrifuging, performing inoculated culture, performing passage, and finally performing resuspension and freeze-drying by using water for injection to obtain freeze-dried powder. The culture medium is composed of a basic culture medium and a growth additive, and all the components are described in detail. The method can efficiently enrich the specific active product of the cornu cervi pantotrichum wax sheet. The invention further relates to application of the high-activity product obtained based on the cornu cervi pantotrichum stem cells in skin care products (such as water aqua and emulsion) and health care products (such as nutritional supplements and functional drinks), provides a new way for development of related products, and has potential economic and social values.

The invention discloses a pig chimeric embryo culture medium and application thereof in effectively improving the in-vitro chimeric ability of pig embryonic stem cells, and belongs to the technical field of bioengineering. According to the pig chimeric embryo culture medium disclosed by the invention, a neutral culture environment for promoting synchronous development of stem cells and embryos is created by accurately regulating and controlling growth factors, nutrient substances and other key components in the culture medium. On the basis, overexpression of anti-apoptosis factors, optimization of microinjection conditions and adoption of a laser-assisted incubation means are combined, so that the chimeric efficiency is further improved, integration of the stem cells and embryos is effectively supported, the chimeric efficiency is improved, and a reliable in-vitro tool is provided for evaluating the chimeric ability and pluripotency of the pig stem cells.

The invention discloses a polypeptide for promoting proliferation of human umbilical cord mesenchymal stem cells and application of the polypeptide in in-vitro culture of the human umbilical cord mesenchymal stem cells. The mesenchymal stem cells have all characteristics of stem cells, have self-renewal and multidirectional differentiation potentials, and can be differentiated into various mesenchymal tissues, such as bones, cartilages, fat and the like. How to rapidly proliferate the human umbilical cord mesenchymal stem cells in vitro so as to rapidly obtain enough seed cells in a short time has important significance. The invention provides a polypeptide with a sequence as shown in Sequence NO.1. The polypeptide has an obvious effect of promoting in-vitro proliferation of human umbilical cord mesenchymal stem cells, does not change phenotypes of the human umbilical cord mesenchymal stem cells, and can be used for in-vitro efficient amplification of the human umbilical cord mesenchymal stem cells.

The invention provides a porcine oocyte in-vitro maturation culture solution additive and application thereof, and belongs to the field of embryo engineering, and the additive contains taurine. The additive also contains sodium pyruvate, cysteine, glucose, NaHCO3, fetal calf serum, estradiol, pregnant horse serum gonadotropin and human chorionic gonadotropin. When the porcine oocyte in-vitro maturation culture solution added with the additive is used for culturing porcine oocytes in vitro, the ROS content in the porcine oocytes can be remarkably reduced, the oxidation resistance of the porcine oocytes can be improved, and the mitochondrial distribution of the oocytes can be improved, so that the development of the oocytes can be improved; the maturation of porcine oocytes and the development of early embryos are promoted. When the technology is applied to the pig assisted reproduction technology, the success rate and quality of embryo engineering can be improved; the pig oocyte in-vitro maturation culture solution added with the additive can improve the premature embryo division rate in parthenogenetic activation of the pig oocytes.

The invention discloses a sebastes schlegeli testis cell line as well as a construction method and application thereof, and belongs to the technical field of marine fish cell culture. Primary culture and subculture are carried out on sebastes schlegeli juvenile fish testis tissues, so that the sebastes schlegeli testis cell line capable of being continuously subcultured in vitro for a long time is established. The sebastes schlegeli testis cell line established by the invention is in a fibroblast shape, has a good growth curve, and has relatively strong multiplication capacity. The lentiviral vector can significantly express the stromal cell marker gene temp2, can transfect the cell line, and can be subsequently applied to research of gonad development related functional genes. At present, the cell line has been subjected to in-vitro continuous passage for more than 60 generations, can be subjected to cryopreservation, and is high in recovery survival rate. Establishment of the cell line provides a stable and excellent experimental material for research of fish gonad development genes.

The invention relates to the technical field of NK cell culture, and discloses a serum-free medium for NK cells from umbilical cord blood after cryopreservation and a culture method thereof.The medium comprises a coating solution, an induction medium and an amplification medium; wherein the coating liquid is prepared from an anti-CD16 monoclonal antibody and DPBS (diphenylphosphonic acid); the induction culture medium comprises a basic culture medium, a human-derived platelet lysate and cell factors IL-15, IL-18 and IL-21; the amplification culture medium comprises a basic culture medium, a human-derived platelet lysate and IL-2. The culture method comprises the following steps: obtaining mononuclear cells from fresh umbilical cord blood, and then cryopreserving and resuscitating; and carrying out multiplication culture on the recovered umbilical cord blood mononuclear cells by using the serum-free culture medium. According to the NK cell culture medium, the NK cells with high purity, high killing activity and high cytotoxic activity can be obtained without autoserum culture.

Cell culture media comprising antioxidants are provided herein as are methods of using the media for cell culturing and polypeptide production from cells. Compositions comprising polypeptides, such as therapeutic polypeptides. produced by the methods herein are also provided.

The invention belongs to the technical field of biological medicine, and particularly relates to a prostate cancer organoid culture medium, a preparation method thereof, a prostate cancer organoid and a culture method thereof. The prostate cancer organ culture medium provided by the invention comprises a basic culture medium and a culture additive, and the culture additive comprises FGF-10 (Fibroblast Growth Factor), Zeaxanthin dipalmitate and Polydatin. In the prostate cancer organoid culture medium provided by the invention, the success rate of organoid culture is greatly improved by utilizing the synergistic effect of the FGF-10, the Zeaxanthin dipalmitate and the Polydatin, the cultured prostate cancer organoid is high in growth speed, the passage frequency is remarkably improved, in-vitro culture can be specifically carried out on prostate cancer tissues of people, and the prostate cancer organoid culture medium has the advantages that the prostate cancer tissue culture efficiency is improved; the survival rate and the formation efficiency of the prostatic cancer organs are further improved.

The invention discloses a preparation method of NK (Natural Killer) cells. The method comprises the following steps: (1) separating and purifying the NK cells; (2) primarily amplifying the NK cells in a low-oxygen environment; (3) co-culturing the NK cells; and (4) continuously amplifying and harvesting the NK cells. The NK cell prepared by the method shows relatively high cell purity and activity, and shows a remarkable anti-tumor effect in a malignant tumor treatment mouse model. The method has the advantages of high efficiency and reliability, and provides good reference for research and clinical application of the NK cells.

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 relates to a culture medium and a culture method of persistent DC-CIK cells and application of the culture medium and the culture method, and belongs to the technical field of cell culture. The invention provides a culture medium for persistent DC-CIK cells, which takes a serum-free culture medium as a solvent and comprises a recombinant human granulocyte macrophage colony stimulating factor, a recombinant human interleukin-12, a recombinant human tumor necrosis factor-alpha, a recombinant human interferon-gamma, L-cysteine, glutathione, ginseng polysaccharide and streptomycin sulfate. According to the culture medium and the culture method of the persistent DC-CIK cells, proliferation and growth of the DC-CIK cells can be promoted, and very high tumor killing activity of the DC-CIK cells can be maintained for a long time.

The invention provides a bone organ model as well as a construction method and application thereof, and the method comprises the following steps: arranging a biological material with a micropore array in a cell culture plate to obtain a bone organ chip; suspending the initial stem cells in a culture medium, and promoting the cells to form a spherical structure through cell aggregation or by using a non-adhesive culture surface, so as to obtain cell spheres; and culturing the cell spheres in the bone organ chip by adopting the culture medium for constructing the bone organ model to obtain the bone organ model. The skeletal organ is successfully constructed through in-vitro self-organized 3D co-culture initial stem cells including embryonic stem cells, induced pluripotent stem cells, bone marrow mesenchymal stem cells, osteoblasts, bone cells, all levels of osteogenic precursor cells, osteogenic-like cells and MC3T3-E1 cells, and the skeletal organ can become an important tool for researching and treating bone related 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 culture medium for culturing a liver cancer micro-tumor model and a culture method thereof. The invention relates to the technical field of biology, and provides a culture medium for culturing a liver cancer micro-tumor model, and the culture medium is composed of an antibacterial and antifungal agent tri-antibody, HEPES, GlutaMax, a human recombinant protein EGF, FGF-10, HGF, IL-2, IL-15, R-spondin 1, calomel, gastrin, A83-01, N-acetyl-L-cysteine, nicotine, an N2 additive, B27 and a basal culture medium. The liver cancer micro-tumor model obtained by the method can accurately reflect various characteristics of the original focus of a patient, and is a good scientific research experiment model and a preclinical experiment model in the field of precise tumor diagnosis and treatment. It can be foreseen that the culture method has wide application prospects in the fields of liver cancer research and clinical diagnosis and treatment.