113 results about "Matrigel" patented technology

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

The invention discloses a carbon dot, hydrogel containing the carbon dot as well as a preparation method and application of the hydrogel, and relates to the technical field of carbon materials. The carbon dot is prepared from the following raw materials: a hydroxyl-containing flavonoid compound and epsilon-polylysine; the carbon dots contain aniline bonds formed by hydroxyl groups of the hydroxyl-containing flavonoid compounds and amino groups of the epsilon-polylysine. The carbon dot shows higher tumor cell killing property and lower normal cell toxicity than the raw material hydroxyl-containing flavonoid compound at the cellular level, can obviously induce immunogenic death of cancer cells and does not induce up-regulation of extracellular matrix collagen, and also has a better effect of regulating a tumor immune microenvironment.

The invention discloses a construction method and application of an animal model for evaluating cardiotoxicity of antitumor drugs, sodium carboxymethyl cellulose (CMC-Na) with extremely low cost is adopted to replace matrigel, a tumor-bearing mouse model with high stability can be constructed at low cost, and experimental verification shows that 4T1 cells are resuspended by using a 0.7% CMC-Na solution, and the tumor formation rate of subcutaneous inoculation reaches gt; 95%. The tumor microenvironment-mediated cardiac injury critical period is accurately captured, and the constructed tumor-bearing mouse model is the optimal tumor-bearing mouse model for evaluating the cardiotoxicity of an antitumor drug (adriamycin) in the tumor microenvironment in the third week after the mouse has a 50mm < 3 > tumor.

The invention belongs to the technical field of bioengineering, and relates to heparan sulfate defect type matrigel as well as a preparation method and application thereof. The preparation method comprises the following steps: providing natural matrigel; providing immobilized heparinase, wherein the immobilized heparinase can specifically catalyze and degrade heparan sulfate in the natural matrigel; carrying out contact reaction on the natural matrigel and the immobilized heparinase; and after the reaction is finished, separating the immobilized heparinase. According to the preparation method, the characteristics that the immobilized enzyme is convenient to separate and recycle are utilized, the problems that in a traditional solution enzyme method, enzyme residues exist, the reaction is difficult to accurately control, and subsequent purification steps are complex are solved, and a standardized heparan sulfate defective matrigel product which is controllable in component, free of residual enzyme and high in stability is provided. The method has wide application value in the fields of tumor mechanism research, anti-tumor drug screening, tissue engineering and the like.

The invention relates to the technical field of biology, and discloses a construction method of a gastric cancer organoid culture system, which comprises the following steps: preparation of a conditioned medium: culturing L-WRN cells and collecting the conditioned medium; preparing a gastric cancer organoid culture medium, and mixing the conditioned culture medium, the basic culture medium, a plurality of growth factors and additives; and extraction and culture of organoid: separating cells from gastric cancer tissues, mixing the cells with matrigel, and culturing by using the prepared culture medium. According to the method, the repeatability and the stability of the organoid model are improved, the response consistency of the organoid in drug screening is improved, and the reliability of the model in disease mechanism research and personalized medical application is guaranteed.

This invention relates to a method for constructing a mouse orthotopic model of highly aggregated glioblastoma, belonging to the fields of biomedicine and experimental animal model technology. Glioblastoma cells are resuspended in a composite carrier of a specific ratio of matrix gel and serum-free culture medium, and a low-temperature injection technique is used. Utilizing the thermosensitivity of matrix gel in the mouse intracranial environment (i.e., its phase transition from liquid to gel at a certain temperature), precise colonization and physical locking of tumor cells at target coordinate points are achieved. This method improves upon existing glioblastoma mouse orthotopic model construction techniques that suffer from tumor cell diffusion and loss, low tumor formation rate, irregular tumor morphology, large intragroup variability, and needle reflux due to low viscosity of the inoculation carrier. It provides a good disease model for subsequent screening or evaluation of anti-glioblastoma drugs.

The present invention relates to a preparation method of a composition for culturing lung organoids, a composition thereby, and an organoid culture method using the same. The present invention is capable of creating an environment that is more similar to an actual tissue than a conventional MATRIGEL®-based culture system, and in particular, exhibits an effect of facilitating tissue differentiation in lung organoid culture and effectively developing into a form that is similar to an actual tissue.

Disclosed is a chemically defined airway basal stem cell culture medium. The provided culture medium involves a simple preparation method and high safety, and does not require a feeder layer or a matrigel-coated culture plate. By means of the culture medium, primary cultured airway basal stem cells achieve good stability; and after multiple passages, the normal morphology of the basal stem cells can be maintained and good differentiation potential can be maintained. In addition, the culture medium has clear components, and can meet both scientific research-grade and clinical-grade applications of airway basal stem cells.

The invention relates to the technical field of biology, in particular to a method for efficiently obtaining human definitive endoderm cells in vitro, which comprises the following steps: S1, inoculating human pluripotent stem cells into matrigel, and performing maintenance culture for 16-24 hours by using a stem cell maintenance culture medium; s2, from Day-2 to Day-1, absorbing and abandoning the stem cell maintenance culture medium, adding a pretreatment culture medium for culture, and changing the pretreatment culture medium once every day; the pretreatment culture medium is a stem cell maintenance culture medium added with dimethyl sulfoxide; s3, carrying out Day0 to Day2, absorbing and abandoning the pretreatment culture medium, adding a definitive endoderm differential culture medium, carrying out induced culture, and changing the definitive endoderm differential culture medium once a day; the definitive endoderm differential culture medium is an RPMI1640 culture medium added with fetal calf serum and activin A; s4, Day3, obtaining the required human definitive endoderm cells, and solving the problems that the existing definitive endoderm differentiation efficiency and cell population purity are limited and are influenced by batch-to-batch variation.

This invention belongs to the field of biomedical technology, specifically relating to a culture reagent composition and method for organoids from hematologic malignancies. The invention first provides a reagent composition for culturing organoids from hematologic malignancies, including acute lymphoblastic leukemia or acute myeloid leukemia; the reagent composition includes a bone marrow microenvironment mimicking matrix reagent and a specific culture medium. Using the reagent composition of this invention, hematologic malignancies can be cultured efficiently with high survival rates. Furthermore, the bone marrow microenvironment mimicking matrix component used in this invention, compared to the commonly used Matrigel matrix in the field, allows for more accurate phenotypic differentiation of hematologic malignancies.

This invention discloses an intestinal fungus Saccharomyces cerevisiae M7015 and its applications belong to the field of microbial technology. Saccharomyces cerevisiae The accession number for M7015 is GDMCC No. 66195. This invention first demonstrated, using a mouse subcutaneous stroma embolism model, that the aforementioned strain can promote angiogenesis; secondly, using mouse and Bama miniature pig myocardial infarction models, it was found that the aforementioned strain can promote collateral angiogenesis in ischemic areas of the heart and improve cardiac function. This invention not only fully elucidates... Saccharomyces cerevisiae M7015 promotes the formation of collateral vessels in the heart and improves cardiac function, and also reveals the broad application prospects of this bacterium in the treatment of vascular diseases.

The invention discloses a construction method and application of a cerebellar organoid model, and belongs to the technical field of organoid culture. According to the method, the stability and repeatability of cerebellar organs are remarkably improved by optimizing the components of the culture medium and the induction process. Key morphogenetic factors are regulated and controlled in a stepped mode, and cells are accurately guided to be directionally differentiated to cerebellum; meanwhile, a culture medium without Matrigel is used in the nerve expansion and maturation stages, batch variation of exogenous matrixes is eliminated, and development path consistency and cell composition uniformity are ensured. The organoid constructed on the basis of the method has remarkable advantages in SCA3 disease modeling, single cell sequencing proves that the difference in batches is remarkably smaller than that of an existing method, and a reliable in-vitro human model platform is provided for SCA3 mechanism research and drug screening.

The invention discloses a method for large-scale cryopreservation of animal cells. The method comprises the following steps: carrying out first-stage amplification on to-be-preserved cells under the support of matrigel; performing second-stage amplification on the to-be-preserved cells subjected to the first-stage amplification under the support of a microcarrier; mixing the to-be-preserved cells with a serum-free cryopreservation solution to prepare a micro-capsule suspension; and putting the microcapsule suspension into a cryopreservation container to carry out cryopreservation. According to the invention, through amplification of two stages, the to-be-preserved cells have reached a high density of 109 orders of magnitude before cryopreservation, a re-amplification link in a cell recovery stage can be greatly shortened, and the to-be-preserved cells are subjected to microencapsulation through the micro-fluidic chip and then cryopreserved, so that physical protection is given to the to-be-preserved cells; the damage of ice crystals in the cryopreservation process is reduced, the survival rate after resuscitation is improved, and the microencapsulated cells can also be directly subcultured, so that the resuscitation operation is simplified; according to the cell cryopreservation device, the cryopreservation container with a large volume is used, so that the cost of cell cryopreservation is reduced.

The invention provides a whole-brain organoid based on human induced pluripotent stem cells as well as a construction method and application thereof, and belongs to the technical field of organoid construction. The method comprises the following steps: culturing human induced pluripotent stem cells in a complete culture medium to obtain a cell aggregate; culturing in an embryoid body forming culture medium to obtain a spherical embryoid body; culturing in a nerve induction culture medium tissue to form a nerve epithelium structure; embedding the single embryoid body with the nerve epithelium structure in a Matrigel gel drop, culturing in a nerve expansion culture medium to form a nerve epithelium-like structure, and replacing a mature culture medium for culturing to obtain the whole-brain organ. According to the method, by dynamically and accurately regulating and controlling culture parameters, the control ability of a cell differentiation area is improved, and interference of non-target cell types is reduced, so that the efficiency and consistency of embryoid formation and neural differentiation are directly improved, and finally, the obtained mature organoid has a more complete epithelial structure and cell polarity.

The application discloses application of a composite hydrogel in promoting cell spheroid culture. The composite hydrogel is prepared from crosslinking of polyvinyl alcohol and sodium alginate. In the cell spheroid culture, a polyvinyl alcohol / sodium alginate composite hydrogel pad is arranged at the bottom of a culture solution, and cells are cultured above the polyvinyl alcohol / sodium alginate composite hydrogel. The method can make the cultured cells quickly form spheroids, and has obvious advantages compared with a currently most widely used matrix gel in the market. In addition, the technology uses polyvinyl alcohol and sodium alginate as main raw materials, both of which are mature commercial polymers, and the cost is low. Moreover, the preparation method through calcium ion crosslinking is simple and efficient.

The invention discloses an ovarian cancer organoid-macrophage co-culture system as well as a preparation method and application thereof. The preparation method comprises the following steps: providing ovarian cancer cells; the method comprises the following steps: resuspending ovarian cancer cells in matrigel, then inoculating the matrigel on which the ovarian cancer cells are resuspended into a culture container, after the matrigel is completely solidified, adding the matrigel into an ovarian cancer organoid complete culture medium, culturing to obtain an ovarian cancer organoid wrapped in the matrigel, and separating to obtain the ovarian cancer organoid; providing macrophage M2, mixing the macrophage M2 and the ovarian cancer organoid, and culturing by using a co-culture medium. In combination with a specific embodiment, according to the preparation method of the ovarian cancer organoid-macrophage co-culture system, macrophages M2 and ovarian cancer organoid are mixed and then cultured with a co-culture medium, the obtained ovarian cancer organoid-macrophage co-culture system contains the macrophages M2, and the functions of the macrophages can be researched.

The invention discloses a method for preparing a vascular organ chip from human umbilical artery stem cells, and belongs to the field of biomedical engineering and tissue engineering. The method comprises the following steps: separating and culturing human umbilical artery stem cells to the third generation; preparing a PDMS micro-fluidic chip containing a liquid inlet channel, a liquid outlet channel, a growth channel and a central culture chamber; a blood vessel precursor aggregate is obtained through cell digestion, aggregate construction, mesoderm induction and blood vessel pedigree induction, the blood vessel precursor aggregate is inoculated into a Coll-1-Matrigel gel modified chip through a two-step method, and dynamic perfusion culture is conducted in combination with a culture medium containing VEGF-A and FGF-2. And finally, the structure and the function of the vascular organ are verified through immunofluorescent staining and fluorescent microbead perfusion. The problems that in the prior art, cell sources are limited, culture conditions are single and the like are solved, the high angiogenesis potential of the human umbilical artery stem cells and a dynamic culture system are utilized to construct a high-bionic-degree vascular network, and the method can be applied to drug screening, vascular pathological model construction and regenerative medicine research.

The invention discloses a semisolid culture medium for screening hybridoma cells. The semisolid culture medium comprises a basic culture medium, a three-dimensional skeleton component and a functional active component, wherein the three-dimensional skeleton component comprises enzymolysis matrigel and methyl cellulose. According to the invention, a brand new combination of synergistic effect of a matrigel-methylcellulose three-dimensional skeleton and a hyaluronic acid-heparin sodium functional compound system is constructed. The culture medium can significantly improve clone integrity, promote rapid proliferation of cells and efficiently enrich high-expression clone, realizes breakthrough of mainstream commercial culture media in screening efficiency and quality of monoclonal hybridoma cells, and is suitable for preparation of monoclonal antibodies.

The invention discloses an organoid ball transfer system, a transfer method and an automatic organoid culture platform, and relates to the field of organoid culture. The invention aims to solve the problems that the existing culture and transfer planting of the organoids mainly depend on manual operation, the transfer efficiency, the position precision and the culture consistency are difficult to guarantee, and the stability and the repeatability of experimental results are influenced by the manual operation. According to the organ-like ball transfer system, the transfer platform is adopted to execute the transfer work of the organ-like balls; and adopting a visual processing unit to track and position the organ-like ball in the transfer process of the organ-like ball. The organoid cell balls can be quickly and nondestructively transferred into the liquid matrigel, and the blank of an organoid automatic system of a small chamber culture system is filled.

The application belongs to the technical field of bioengineering, and relates to a heparan sulfate-deficient Matrigel as well as a preparation method and application thereof. The preparation method comprises the following steps: providing natural Matrigel; providing immobilized heparinase which can specifically catalyze degradation of heparan sulfate in the natural Matrigel; contacting and reacting the natural Matrigel with the immobilized heparinase; and separating the immobilized heparinase after the reaction is completed. The preparation method utilizes the characteristics of the immobilized enzyme that is convenient to separate and recover, overcomes problems such as enzyme residue, difficulty in accurately controlling the reaction and complexity of subsequent purification steps in the traditional solution enzyme method, and provides a standardized heparan sulfate-deficient Matrigel product which is controllable in composition, free of residual enzyme and high in stability. The application has wide application value in the fields of tumor mechanism research, anti-tumor drug screening and tissue engineering.

The invention relates to the technical field of biological medicine, in particular to a construction method and application of a respiratory virus infected alveolar organ model. Comprising the following steps: infecting alveolar cell source single cells by using a virus inoculum, coating the infected alveolar cell source single cells with matrigel, and inoculating the infected alveolar cell source single cells into a first culture medium for three-dimensional culture to obtain the respiratory virus infected alveolar organ model. According to the invention, alveolar cells are creatively applied to model characterization of respiratory virus infection, and particularly, alveolar cell source single cells are subjected to infection treatment by adopting influenza A H1N1 virus, so that the constructed model can well simulate and characterize the infection process of influenza A virus in human lung organs; the method provides an experimental basis for drug research and development (such as high-throughput screening of anti-influenza virus drugs), viral disease pathogenesis research, research on defense mechanisms of naturally occurring virus infection or virus infection caused by experiments and the like, and has important guiding significance.

The application discloses a method for culturing NK cells and application thereof. The application can harvest high proportion and more NK cells by rotating culture of hPSCs in a 3D culture device to form EBs spontaneously, and then further inducing differentiation of the NK cells by using mixed cellulose ester (MCE). The method of the application does not depend on 96-well plates, special consumables or matrigel and the like matrix glue, has high sphere formation efficiency and is convenient to operate, and has wide application prospect.

The invention relates to the technical field of cytobiology and regenerative medicine, and discloses a composition and method for efficiently and directionally inducing iPSC (induced pluripotent stem cells) to be differentiated into functional microglial cells, and the method comprises the following steps: S1, digesting the induced pluripotent stem cells into single cells, resuspending the single cells by using an embryoid induction culture medium, inoculating the single cells into a low-adsorption culture plate, and culturing for 2-4 days to obtain a culture medium; forming an embryoid; s2, transferring the obtained embryoid into a matrigel-coated culture plate, adding a myeloid precursor cell induction culture medium for culturing, regularly changing the liquid during the period, and culturing for 2-3 weeks to obtain suspended myeloid precursor cells; s3, collecting the obtained myeloid precursor cells, centrifuging the myeloid precursor cells, re-suspending the myeloid precursor cells by using a microglial cell end induction culture medium, inoculating the myeloid precursor cells into a culture plate, adding the microglial cell end induction culture medium for culturing, regularly changing the medium during the culture period, and culturing for 10-14 days to obtain mature microglial cells. The invention solves the problems of long period and high cost of the existing scheme for differentiating the iPSC into the microglial cells.

The application relates to a tumor organoid-immune cell co-culture method, a co-culture system and application thereof. The method comprises the following steps: S1. adding pre-cooled mixed matrix material into a culture container, and forming a matrix layer after solidification; wherein the mixed matrix material comprises Matrigel matrix glue and gelatin hydrogel in a volume ratio of 1: (0.1-10); S2. inoculating tumor organoid suspension obtained through pretreatment onto the surface of the matrix layer, so that the tumor organoid is attached to the matrix layer, and dropping immune cell suspension obtained through pretreatment onto the surface of the tumor organoid attached to the matrix layer; and S3. adding co-culture medium into the culture container, so that the tumor organoid and the immune cell are immersed in the co-culture medium, and dynamic co-culture is carried out. The method solves the problems of poor co-culture compatibility, poor stability and low repeatability in the prior art, and realizes long-term, efficient and stable co-culture of the tumor organoid and the immune cell.

The invention discloses a culture medium for goat trophoblast organoid, a culture method and application, and belongs to the field of goat trophoblast organoid culture. The method comprises the following steps: separating trophoblast tissues from a healthy goat placenta which is pregnant for 80-90 days, digesting the trophoblast tissues through type II collagenase to obtain primary cells, compounding the primary cells with matrigel, and performing three-dimensional culture in an improved trophoblast organ culture medium with specific components to form mature organs. The organ is of a typical three-dimensional structure, and KRT23 and SOX2 markers are stably expressed. The organoid culture system based on the goat primary trophoblast cells is established for the first time, gene modification is not needed, the in-vivo physiological status is highly simulated, and the culture system is standardized and good in repeatability. The constructed organoid can be used as an ideal in-vitro model for researching placenta development, evaluating drugs and toxins and modeling infectious diseases in the gestation period, and is of great significance in improving the reproductive research level of goats and the production benefits of animal husbandry.

The invention discloses an ovarian cancer organoid-T cell co-culture system as well as a preparation method and application thereof. The preparation method comprises the following steps: providing ovarian cancer cells; the method comprises the following steps: resuspending ovarian cancer cells in matrigel, then inoculating the matrigel on which the ovarian cancer cells are resuspended into a culture container, after the matrigel is completely solidified, adding the matrigel into an ovarian cancer organoid complete culture medium, culturing to obtain an ovarian cancer organoid wrapped in the matrigel, and separating to obtain the ovarian cancer organoid; t cells are provided, and the T cells and the ovarian cancer organoid are mixed and then cultured with a co-culture medium. In combination with a specific embodiment, according to the preparation method of the ovarian cancer organoid-T cell co-culture system, T cells and ovarian cancer organoid are mixed and then cultured by a co-culture medium, the obtained ovarian cancer organoid-T cell co-culture system contains the T cells, and T cell immune response can be truly reflected.

The invention provides a preparation method of SPF (specific pathogen free) pigskin extracellular matrix (ECM) hydrogel. The preparation method comprises the following steps: S1, collecting SPF pigskin and pretreating; s2, the pretreated SPF pigskin is subjected to degreasing treatment; s3, sterilizing and disinfecting the freeze-dried powder, and then carrying out enzymolysis; s4, adjusting the pH and osmotic pressure of supernate after enzymolysis to obtain a hydrogel product. The source of the hydrogel product does not contain non-biological source materials at all, establishment of a three-dimensional cell culture environment and growth of cultured cells, especially organoids are facilitated, and meanwhile the problems of tumor sources, batch differences, high cost, potential safety hazards and the like of existing hydrogel products such as Matrigel and the like are solved. Experimental results show that the pigskin ECM hydrogel with different concentrations and from different ages can support the growth of intestinal organs, and the number of the organs cultured by the SPF pigskin ECM hydrogel with the concentration of 3 mg / mL is even more than that of Matrigel.

The invention relates to the field of aerospace biology and cell biology, in particular to an in-vitro preparation method of myocardial cells under a supergravity condition. Comprising the following steps: step 1, performing adherent culture on myocardial cells on a two-dimensional matrix; 2, mixing the myocardial cells subjected to two-dimensional culture in the step 1 with matrigel to form a three-dimensional culture system; step 3, placing the three-dimensional culture system obtained in the step 2 in a supergravity environment for culture; wherein the gravity intensity of the supergravity environment is 3G, and the treatment time is 12-96 hours. The invention provides a complete and repeatable myocardial cell in-vitro preparation method under the supergravity condition for the first time. A stable 3G supergravity environment can be provided, the damage of high shear force to cells is effectively reduced, the core pain point that a traditional supergravity experiment depends on expensive and rare parabola flight or a large centrifugal machine is solved, and the research cost and the technical threshold are greatly reduced.

The present application provides a method for in vitro purified corneal limbal stem cells to be transdifferentiated into lens spherules, specifically, the present application inoculates corneal limbal stem cells onto a culture plate containing a Matrigel glue layer, and cultures the corneal limbal stem cells in a culture system containing Y-27632 additives in LSCM culture solution, so that lens spherule organoids can be obtained. The method of the present application can realize the rapid transdifferentiation of corneal limbal stem cells into lens spherules, so that they can be implanted into the capsular bag as seed cells for in situ regeneration of the lens.

The invention provides an umbilical cord matrigel stem cell culture medium as well as a preparation method and application thereof, and belongs to the technical field of stem cells. Comprising the following raw materials: polyethylene glycol, cane sugar, an exosome composition, a cell factor composition, globulin, tween-80 and a PBS buffer solution. The culture medium can promote the growth of cell mitosis and mesoderm cells, the cells are not prone to deformation in in-vitro culture, the cell clone formation rate retention rate after multiple passage is high, the cells can still keep the characteristics of primary cells after being gradually differentiated, and the culture medium has wide application prospects.