10731results about "Cell culture active agents" patented technology

The invention relates to a high-activity recombinant XVII type collagen as well as a rapid preparation method and application thereof, and belongs to the technical field of bioengineering. The amino acid sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 1, and the nucleotide sequence of the high-activity recombinant XVII type collagen is as shown in SEQ ID NO. 2. The invention also provides a preparation method and application of the recombinant XVII type collagen. The high-activity recombinant XVII type collagen has good biological activity, has good effects of promoting cell adhesion, cell proliferation and cell migration, and can be compounded with other proteins to form raw materials of tissue engineering products, cosmetics, health care products or medicines. The high-activity recombinant XVII type collagen gene is integrated to different positions of a yeast genome through three different vectors, and the copy number of a target gene is increased, so that the expression quantity of the recombinant XVII type collagen is remarkably increased, and the yield of the high-activity recombinant XVII type collagen is effectively increased.

The invention belongs to the technical field of biology, and discloses a reagent combination or kit for constructing intestinal organs and application of the reagent combination or kit. According to the reagent combination or the kit, intestinal organs can be obtained from cell-derived epithelial cells obtained from a donor in a non-invasive manner, and the obtained intestinal organs can be cryopreserved and recovered and can be amplified in vitro for a long time; transcriptome characteristics are similar to those of real human small intestine tissues, and typical marker genes of various small intestine pedigree cell types are highly expressed; compared with intestinal organs obtained through induction of pluripotent stem cells, the intestinal organs have more intestinal pedigree characteristics, and the intestinal function development is more mature; after being promoted to be mature, the intestinal organ also highly expresses genes related to drug absorption and metabolism, has drug absorption capability similar to that of an immortalized intestinal cell line, but more prominently shows intestinal cell lineage characteristics, is closer to an intestinal environment in a real human body, and can be used for screening intestinal disease drugs; the intestinal barrier function is realized.

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.

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 belongs to the technical field of protein engineering and biology, and discloses a recombinant III-type humanized collagen, a coding gene thereof, a protein preparation, a preparation method of the protein preparation, application of the protein preparation and a skin care product. The amino acid sequence of the recombinant III type humanized collagen is as shown in SEQ ID NO. 1. According to the recombinant III type humanized collagen, the transdermal absorption efficiency is greatly improved, and meanwhile, the proliferation and migration capabilities of keratinocytes are also remarkably improved. The collagen is widely applied to the fields of medical cosmetic plastic surgery, cosmetics and the like, and has a good application prospect.

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 discloses a method and an index for in-vitro prejudgment of an in-vivo immunoregulation relationship of algal polysaccharides, and researches show that algal polysaccharides CRVP can regulate interaction between macrophages and liver cancer cells. The CRVP promotes macrophages to be polarized into M1 type, plays a proinflammatory role, further affects the migration ability of the liver cancer cells, promotes the liver cancer cells to generate differential metabolites GABA, and induces apoptosis of the liver cancer cells at the same time. In addition, CRVP can also act on liver cancer cells, further influence the proliferation and phagocytic ability of macrophages, and induce the macrophages to be polarized into M1 type.

Disclosed herein are antibodies that specifically bind to Gal3 and methods of use thereof. In some embodiments, also described herein are methods of inducing immune activation or promoting T cell or Natural Killer cell proliferation with an antibody that specifically binds to Gal3. Also disclosed herein are methods and compositions of reducing fibrosis or propensity thereof in a tissue with antibodies that specifically bind to Gal3. In some cases, the anti-Gal3 antibody also disrupts the interaction between Gal3 and TIM-3.

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 the technical field of biological medicines, and particularly discloses an anti-human CD3 rabbit recombinant monoclonal antibody as well as a preparation method and application thereof. The antibody obtains anti-CD3 immune response by immunizing a rabbit, is obtained by combining single B cell sorting, gene cloning and a mammalian cell expression system, and has high affinity and good specificity. The antibody can effectively recognize CD3 positive T cells in human peripheral blood and can be applied to the fields of flow cytometry, immunohistochemistry, T cell function research and the like. Experimental results show that the antibody has the advantages of high titer, strong fluorescence signal, low background and the like, and is superior to the traditional murine anti-CD3 antibody. Amino acid sequences of light chain and heavy chain variable regions of the antibody are further obtained, and controllability and repeatability of antibody expression are achieved. The antibody has important scientific research value and industrial application prospect, and is suitable for various scenes such as T cell marker detection, immune state evaluation, immunotherapy auxiliary research and the like.

The invention relates to a heart organoid and an in-vitro preparation method thereof, and belongs to the technical field of stem cells. The invention provides a heart organoid. The heart organoid is obtained by induced culture of an adherent pluripotent stem cell mass; the adherent pluripotent stem cell mass is obtained by adherent culture of a suspended pluripotent stem cell mass; the suspension pluripotent stem cell mass is obtained by suspension culture of pluripotent stem cells. According to the in-vitro preparation process of the heart organoid, after pluripotent stem cells form balls, the pluripotent stem cell balls are subjected to staged induced differentiation under adherent culture, so that cell subtypes such as epicardial cells and the like can be generated without additional addition or additional stimulation of signal channels, and the heart organoid is more homogeneous. The self-occurring multi-lineage coexisting heart organoid can better simulate biological events derived from multiple cell types and multiple cell subtypes in the early development process of an embryonic heart, and has great application prospects in exploration of pathogenic factors and potential treatment targets of congenital heart diseases.

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 provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1 and application thereof, and relates to the technical field of biology. The invention provides an enhancer RNA (Ribonucleic Acid) molecule MZGAe1. The nucleotide sequence of the enhancer RNA molecule MZGAe1 is as shown in SEQ ID NO: 1; meanwhile, the invention further provides a specific sgRNA sequence for activating the molecule in a targeted manner and a recombinant vector of the specific sgRNA sequence. Expression of endogenous MZGAe1 of cells is specifically activated by adopting a CRISPR activation technology, and conversion of mouse embryonic stem cells to bicellular-like cells can be efficiently promoted. By providing a brand-new endogenous targeted MZGAe1 accurate activation tool, the efficient transformation of wild mouse embryonic stem cells to bicellular cells can be realized only under the condition of endogenous activation of single enhancer RNA molecule MZGAe1, the proportion is at least increased by 5%, the expression of a totipotent marker gene ZSCAN4 is activated, and the expression of the totipotent marker gene ZSCAN4 is promoted. And an efficient and specific brand-new research tool and scheme are provided for researching zygote genome activation, embryonic development early events and cell reprogramming.

The invention discloses a novel application of a small molecule compound BRD4780 in resisting ricin, the small molecule compound BRD4780 has a remarkable effect of resisting ricin toxicity and can be used in development of anti-ricin drugs, a brand new candidate molecule is provided for development of the anti-ricin drugs, the drug research and development blank in the field is filled, and the application of the small molecule compound BRD4780 in resisting ricin is developed. The method has important significance in the aspects of clinical treatment, biological protection and immunotoxin treatment safety optimization, and is wide in application prospect.

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.

The invention belongs to the technical field of biological medicines, and particularly relates to application of a targeted liposome as an active component in preparation of a medicine for treating ischemic brain injury. The targeted lipidosome for treating cerebral arterial thrombosis is prepared by fusing the lipidosome and the platelet-neutrophil aggregate membrane, realizes efficient and specific targeting of an ischemic brain region, effectively reduces brain tissue infarction and damage of a new region, relieves brain inflammation through multi-channel regulation and control, and improves the cerebral arterial thrombosis treatment effect. The compound can be used as an active ingredient for preparing a medicine for treating ischemic brain injury. And after the target liposome is further coated with a rutin drug, the synergistic treatment effect is realized. The liposome medicine can be used as an active component and has a good application prospect in treatment of ischemic brain injury diseases.

Methods for production of platelets from pluripotent stem cells, such as human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) are provided. These methods may be performed without forming embryoid bodies or clusters of pluripotent stem cells, and may be performed without the use of stromal inducer cells. Additionally, the yield and / or purity can be greater than has been reported for prior methods of producing platelets from pluripotent stem cells. Also provided are compositions and pharmaceutical preparations comprising platelets, preferably produced from pluripotent stem cells.

Disclosed herein are compositions and methods related to differentiation of stem cells into pancreatic endocrine cells. In some aspects, the methods provided herein relate to generation of pancreatic β cell, α cell, δ cells, and EC cells in vitro. In some aspects, the disclosure provides pharmaceutical compositions including the cells generated according to the methods disclosed herein, as well as methods of treatment making use thereof.

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 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.

Methods, compositions and devices characterized by in vitro maturation of oocytes. In particular, the present disclosure features methods of inducing oocyte maturation in vitro by co-culturing oocytes of a female subject with an ex vivo composition comprising a plurality of ovarian supporting cells (e.g., granular cells). Additional methods of administering a follicle triggering agent and extracting oocytes from a female subject are also provided. Such methods, compositions, and devices are particularly useful for assisted reproductive technology (ART) procedures.

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 present disclosure relates in some aspects to methods, cells, and compositions for preparing cells and compositions for genetic engineering and cell therapy. Provided in some embodiments are streamlined cell preparation methods, e.g., for isolation, processing, incubation, and genetic engineering of cells and populations of cells. Also provided are cells and compositions produced by the methods and methods of their use. The cells can include immune cells, such as T cells, and generally include a plurality of isolated T cell populations or types. In some aspects, the methods are capable of preparing of a plurality of different cell populations for adoptive therapy using fewer steps and / or resources and / or reduced handling compared with other methods.

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.

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.

Type 2 diabetes (T2D) is a clinical syndrome caused by insufficient insulin secretion for insulin requirements. Described herein are compositions and methods for microphysiological MPS models of disease (MODs) for diabetes. These platforms allow one to compare the effect of chronic β-cell stimulation in the presence and absence of patient specific immune cells in IPSC-derived islets from each group. Additionally, one can reproduce the T2D β-cell phenotype, using islets-on-chips will also be exposed to gluco-lipotoxicity. Likewise, skeletal muscle-on-chips are exposed to patient specific activated immune cells, variable motor neuron innervation and lipids characteristic of T2D.