1885results about "Cell culture supports/coating" patented technology

The invention relates to an organ-like core-shell microsphere as well as a preparation method and application thereof. The preparation method comprises the steps that 1, raw materials are prepared, specifically, shell raw materials and inner core raw materials are prepared, the shell raw materials comprise a photoinitiator and methacrylated hyaluronic acid (HAMA), and the inner core raw materials comprise organoid precursor cell suspension and matrigel; 2) molding: extruding the dispersion phase of the shell raw material wrapping the core raw material to a continuous phase by using a micro-fluidic chip and a high-precision injection pump, cutting the dispersion phase into liquid drops by the continuous phase, and performing illumination curing on the liquid drops to form core-shell microspheres; and (3) incubating, namely culturing the core-shell microspheres in a culture medium, so that the organoid precursor cells in the inner core are developed into organoids. According to the preparation method, rapid forming and curing of the organ-like precursor cells carried by the matrigel are achieved, so that the time of the organ-like precursor cells staying in the oil phase is shortened, the cells can obtain oxygen and nutrient substances, and the cell activity is improved.

The invention relates to the technical field of tumor biomedical engineering and organ chips, in particular to a vascularized gastric cancer organ chip and a preparation method thereof.The chip is composed of an integrated micro-fluidic main body, an annular micro-column array and an optical sealing film, and a central culture cavity is divided into a tumor area and a blood vessel area by micro-columns; the preparation method comprises the following steps: injecting a fibrous protein solution containing human umbilical vein endothelial cells and cancer-related fibroblasts into a vascular region, and adding thrombin for in-situ gelation to form a pre-vascularized network; the method comprises the following steps: mixing a patient-derived gastric cancer organ with a methacrylic acid esterified gelatin pre-polymerized solution, injecting the mixture into a tumor area, and carrying out photo-crosslinking immobilization; culturing for 7-14 days under the dynamic perfusion condition of a mixed culture medium to obtain a three-dimensional gastric cancer organ model containing a capillary network; the method is expected to be used for rapid, low-cost and high-throughput screening of chemotherapy or anti-angiogenesis drugs and accurate prediction of individual curative effects of patients.

The invention provides a micro-fluidic chip capable of culturing oral squamous cell carcinoma organoid and a preparation method thereof, and relates to the technical field of micro-fluidic chips, the chip comprises a four-layer structure: the first layer is a closed layer, the second layer is a culture solution / drug pool layer (containing a concentration gradient injection channel and a slow flow channel), the third layer is a matrix culture pool layer, and the fourth layer is a matrix culture pool layer. The fourth layer is a matrigel inflow layer (containing an S-shaped flow channel); the second layer and the third layer are integrally formed, and all the layers are connected through oxygen plasma welding; the problems that in the prior art, matrigel is prone to shifting, drug gradient regulation and control is poor, operation is tedious, and adaptability is insufficient are solved, the matrigel and organoids can be stably fixed, the drug concentration gradient can be precisely regulated and controlled, the nutrition supply efficiency is enhanced, the structural integration level and operation convenience are improved, and the application prospect is wide. The method is suitable for oral squamous cell carcinoma organoid culture, drug screening and tumor mechanism research.

The invention discloses a humanized osteoporosis mouse model and a construction method thereof, and belongs to the technical field of biotechnology and animal models. The invention aims at the core defects of large species difference, insufficient humanization degree, low pathological simulation degree, unstable phenotype and the like of the existing osteoporosis model. The humanized osteoporosis mouse model is constructed through five core steps of experimental animal pretreatment, collagen-nano hydroxyapatite bionic scaffold preparation and cell pre-implantation, immune-bone metabolism double-derived system construction, progressive osteoporosis induction and model identification and verification. According to the method, double-person origination of immune and bone metabolism systems is realized for the first time, the colonization rate and survival time of human cells are greatly improved, the pathological process of clinical postmenopausal osteoporosis is accurately simulated, the phenotype of the model is stable and uniform, the reactivity matching degree with clinical drugs is high, and the method can be widely applied to research and development of anti-osteoporosis drugs and research of pathological mechanisms.

The invention relates to an aqueous composition comprising thiolated and norbornene-modified cold-water species-derived gelatine. A further aspect of the invention relates to a method to generate a three-dimensional hydrogel object, particularly an in vitro model for disease modelling / drug testing or an implant, comprising providing a composition according to the invention and forming the hydrogel object by bioprinting (extrusion-based or digital light-based (DLP)) or molding / casting to provide the three-dimensional hydrogel object. Also provided are a hydrogel implant obtained by a method according to the invention, and a kit comprising dried thiolated and norbornene-modified cold-water marine species gelatine.

The invention provides 3D tumor cell culture hydrogel as well as a preparation method and application thereof. The 3D tumor cell culture hydrogel is prepared from the following raw materials: polyvinylpyrrolidone and polyvinyl alcohol. The preparation method comprises the following steps: mixing a polyvinylpyrrolidone solution with a polyvinyl alcohol solution, adjusting the pH value, and freezing-unfreezing to obtain the 3D tumor cell culture hydrogel. The 3D tumor cell culture hydrogel provided by the invention does not need to be additionally added with a chemical cross-linking agent, and the material is high in biocompatibility and flexible in modulus adjustment, and can be well applied to a 3D in-vitro culture model of tumor cells.

The invention discloses a construction method and application of an in-vitro callus-like organ, the survival rate of the organ is improved by improving freeze-formed bio-ink, and a stable directional channel structure is further prepared, so that the internal mass transfer problem of the callus-like organ is improved, and the material exchange efficiency is improved. The forming mode of the callus-like organ is simple and effective, the stent material is easy to prepare, and the callus-like organ has mature cross-linking and printing characteristics, is expected to be applied to drug screening research or bone defect repair, is clear in process and simple in composition, and has relatively high operability.

The invention relates to the technical field of synthetic biology, in particular to polypeptide with a triple helix structure, recombinant XII type humanized collagen and application of the recombinant XII type humanized collagen. The recombinant XII type humanized collagen is successfully expressed and prepared, has a triple-helix structure and good biological activity including promotion of cell proliferation activity, cell adhesion activity and inhibition of MMP-1, is used as a biological material derived from a human body, does not generate immunological rejection and anaphylactic reaction when applied to the human body, and has a good application prospect. The composition can be used for medical or non-medical application of a plurality of tissues and organs of a human body, filling, compatibilizing or repairing, and promotion of skin compactness or wrinkle resistance and other scenes.

Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGFβ / Activin / Nodal signaling and BMP signaling are provided. Also provided are methoc and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process.

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 mouse lung organ and fibroblast co-culture method, which comprises the following steps of: performing in-vitro mouse lung organ culture on mouse lung tissue primary epithelial stem cell spheres (pneumospheroids) in a Transwell upper chamber, and after the lung organs in the Transwell upper chamber are differentiated, replacing with a lower chamber paved with mouse lung fibroblasts, therefore, a gas-liquid interface co-culture system of the mouse lung organ (upper chamber) and the fibroblast (lower chamber) is formed, and the hierarchical structure, morphological characteristics and regeneration function of the upper chamber organ and the phenotype, function and related signal path activation level of the lower chamber fibroblast can be dynamically researched.

The invention discloses a culture method of high-activity human urine-derived stem cells, which relates to the technical field of biological medicine, and is technically characterized by comprising the following steps: collecting urine samples of pregnant and lying-in women, coating a culture plate with gelatin, inducing adherence, performing passage amplification, performing immunofluorescence identification, performing function detection and other optimized conditions, and separating and culturing the stem cells. Compared with urine-derived stem cells of normal adults, the stem cells derived from pregnant women have stronger activity, higher dryness and faster multiplication capacity. The invention provides a novel and superior cell source for tissue engineering and regenerative medicine.

Provided are immortalized fish fibroblast cells and food products comprising said fish fibroblast cells.

This invention discloses a method for rapidly constructing a human-like cerebral cortex organ-on-a-chip using bio-3D printing and its applications. It comprises three parts: a microfluidic chip fabrication method, a human-like cerebral cortex hydrogel preparation method, and the printing of the human-like cerebral cortex. The microfluidic chip comprises five layers: a mixed-flow channel layer, a liquid pool layer, a micropore array layer, a human-like cerebral cortex culture layer, and a culture medium recovery layer. The human-like cerebral cortex hydrogel is composed of gelatin, alginate, and hyaluronic acid. The human-like cerebral cortex is directly printed into the microfluidic chip using a suspension bath extrusion printing method, and then encapsulated to obtain the human-like cerebral cortex organ-on-a-chip. This invention overcomes the shortcomings of traditional cell culture by directly constructing a large-scale human-like cerebral cortex with three interconnected layers in situ within the organ-on-a-chip using bio-3D printing. Perfusion culture mimics cerebrospinal fluid circulation, facilitating substance exchange, maintaining cell viability, and inducing cell differentiation, and can be widely used in drug development for brain diseases.

The invention discloses a hydrogel microsphere sorting method, an artificial organ preparation method, a system and a medium. The method comprises the following steps: acquiring a hydrogel microsphere image in the micro-fluidic chip; analyzing the hydrogel microsphere image through a neural network to obtain an analysis result; based on the analysis result, hydrogel microspheres wrapping the target cells are sorted out; the neural network comprises a feature extraction layer and a multi-scale joint output head, and the hydrogel microsphere image is processed by the multi-scale joint output head after being convolved by the feature extraction layer to obtain an analysis result. According to the method, the hydrogel microspheres wrapped with the target cells are accurately identified and sorted by using a droplet microfluidic technology, a standardized artificial organ can be prepared based on the sorted hydrogel microspheres, and the functional maturity and reliability of the artificial organ are remarkably improved.

The invention relates to the technical field of biological materials, in particular to composite hydrogel as well as a preparation method and application thereof. In particular to an ion response type ELPs-SA composite hydrogel constructed based on elastin-like polypeptides (ELPs) and sodium alginate (SA) as well as a preparation method and application of the ion response type ELPs-SA composite hydrogel. According to the hydrogel, elastic-like polypeptides ELP-Vs (such as adhesion promotion, angiogenesis, antibiosis and the like) with biological functions are introduced and react with SA, CaCO3 and gluconolactone (GDL) together, and the non-covalent cross-linked three-dimensional network hydrogel is formed under mild conditions. The obtained hydrogel has excellent mechanical strength (the energy storage modulus is 450-1773 Pa), a proper microporous structure (the pore diameter is 50-100 microns) and good biocompatibility, and can support three-dimensional culture of cells and promote angiogenesis and immunoregulation. The material has a wide application prospect in the biomedical fields of tissue engineering, wound repair, organoid construction and the like.

The invention provides hindbrain organoids and methods of producing such organoids. Said hindbrain organoids may include healthy or healthy and diseased tissue. Also provided are methods of testing therapeutic agents using said hindbrain organoids. The invention also provides culture mediums used for producing (patterning) said hindbrain organoids and kits including the culture medias.

The present invention relates to the new use of an exosome secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and in particular relates to the use of the exosome in the preparation of a drug for treating liver diseases. The exosome is secreted by hepatocyte lineage cells differentiated via the directed induction of embryonic stem cells, and the hepatocyte lineage cells are hepatic progenitor cells and / or mature hepatocyte-like cells.

The invention provides recombinant III-type collagen and application thereof, and relates to the technical field of genetic engineering. According to the invention, the yield of the collagen is improved by optimizing the nucleotide sequence of the target gene and optimizing the culture conditions of pichia pastoris, so that the collagen is efficiently expressed in engineering bacteria. The recombinant engineering bacterium shows strong potential in the aspect of improving the yield of protein with high economic value, and is suitable for large-scale industrial production. The recombinant III-type collagen is high in expression quantity, simple in preparation process and suitable for preparation of biological materials for medical cosmetology, wound repair and the like, and has a wide application prospect.

The invention discloses a method for wrapping stem cells by adopting programmable DNA (deoxyribonucleic acid) hydrogel and application of the method, under isothermal physiological conditions, three-dimensional DNA hydrogel is formed through a hybridization chain reaction of an initiation chain (I) and hairpin structure DNA to realize reversible wrapping and release of the stem cells. The hairpin structure DNA is formed by denaturation-renaturation treatment of at least two DNA monomer building blocks, and the mechanical rigidity of the hydrogel is realized by regulating and controlling the molar ratio of the initiation chain (I) to the total amount of the DNA monomer building blocks; the DNA hydrogel can reversibly wrap and release stem cells, and the wrapped stem cells can maintain activity and dryness and recover normal multiplication capacity after being released. According to the method disclosed by the invention, the activity and dryness of the stem cells are remarkably maintained, obvious immune activation is avoided, and the method provides a brand new tool for in-vitro culture and engineering application of the stem cells and has a wide application prospect.

Provided herein are methods for coating a microparticle, cell, or therapeutic agent with a biomimetic zona pellucida (BZP). Also provided herein are BZP-coated cells prepared using the enclosed methods. The methods for using the BZP-coated microparticle to treat diabetes are also provided.

Provided herein are peptide amphiphiles (PAs) comprising a bioactive peptide, nanofibers displaying the bioactive PAs, and methods of use thereof. The disclosed peptide amphiphiles comprise a hydrophobic tail, a structural peptide segment, a charged peptide segment, and a bioactive IKVAV peptide. The disclosed PAs may be used in cell culture methods and in methods of treating central nervous system injury.

Disclosed herein are methods of producing organoids. In some aspects, the methods involve producing organoids by implanting primary cancer cells such as glioblastoma cells into brain organoids, thereby producing brain cancer organoids. The method can comprise an incubation step in which the brain organoid is incubated with primary cancer cells in a hanging liquid droplet to produce the brain cancer organoid. In some aspects, the brain cancer organoids provide tools for understanding and treating cancers of the nervous system, such as glioblastoma.

The invention relates to an extraction and culture method of urine-derived stem cells, and belongs to the field of cell biology and regenerative medicine. The method comprises the following steps: coating a culture plate with a gelatin solution; collecting clean midstream urine of healthy people and adding antibiotics; centrifugally separating the cell precipitate; washing with a phosphate buffer solution; re-suspending the precipitate by using a urine stem cell complete culture medium for primary culture; and after the cells grow to a certain fusion degree, carrying out passage amplification. The urine stem cell complete medium comprises a basic medium, fetal calf serum, epidermal growth factors, platelet-derived growth factors and basic fibroblast growth factors. By adopting the method disclosed by the invention, the urine-derived stem cells can be efficiently and stably separated from urine, and the obtained cells have high multiplication capacity, good stemness maintaining capacity, multidirectional differentiation potential and immunoregulation function, and can provide an ideal seed cell source for treating diseases such as diabetic erectile dysfunction.

The invention relates to a lung cancer organoid and peripheral blood source immune cell co-culture model and a construction method thereof. Specifically, the invention provides a construction method of a macrophage and tumor organoid co-culture model for evaluating anti-tumor activity, monocytes of autologous or allogeneic peripheral blood of a patient are induced and differentiated into high-purity macrophages in vitro, and the high-purity macrophages and lung cancer tumor organoid are subjected to three-dimensional co-culture in matrigel; the model aims at highly reducing a core interaction network of tumor cells and macrophages in TME; the dynamic change of the polarization state of the macrophage is simulated and observed; the problem that an organ-like model is incomplete due to immune component deficiency or spatial positioning distortion of an existing model is effectively solved, an experimental platform closer to the physiological state is provided for tumor immune microenvironment research, and therefore development of the tumor immune treatment field is promoted.

The present disclosure relates to organoids and particularly to brain organoid models. The brain organoids include neurons, microglia, astrocytes, and blood vessels. The brain organoid models can be used to create models for Alzheimer's Disease. Methods of using the brain organoids for drug discovery are also described.

The invention discloses a medicine screening method based on colorectal cancer organoid, which comprises the following steps: adding a colorectal cancer culture medium into the cultured colorectal cancer organoid, culturing for 48-72 hours, adding a to-be-detected colorectal medicine, continuously culturing for 72-96 hours, detecting the cell activity, and screening the medicine capable of reducing the cell activity. According to the method, the tumor microenvironment of the organoid is utilized, the modeling period is shortened, the cost is reduced, the tumor state can be simulated, high-throughput drug screening can be carried out in vitro, and the method has important significance on accurate treatment of lung cancer.

The invention relates to the technical field of biology, in particular to a composition, a culture medium and application of the culture medium in intestinal organ culture. The glucosamine hydrochloride is applied to culture of intestinal organs for the first time, organ proliferation and differentiation can be effectively promoted, and the number of formed organs is remarkably increased. On the basis, a three-dimensional culture system for staged time sequence induction in a postpartum newborn period, a child period and an adult period is constructed, and accurate construction of intestinal organs of the mice in different development periods is realized by combining dynamic regulation and control of an extracellular matrix and combinatorial optimization of growth factors; and an in-vitro model covering multiple time sequence stages is provided for intestinal development mechanism research and disease modeling.

The present invention aims to provide a method for efficiently producing a cell mass containing pituitary tissue from pluripotent stem cells. A method for producing a cell mass containing pituitary tissue, including the following steps (1) and (2):(1) a first step of suspension-culturing pluripotent stem cells to form a cell aggregate in the presence of a Wnt signal transduction pathway inhibiting substance,(2) a second step of suspension-culturing the aggregate obtained in the first step in the presence of a BMP signal transduction pathway activating substance and a Sonic hedgehog signal transduction pathway activating substance, thereby obtaining a cell mass comprising pituitary tissue.

The present application relates to a kind of RIC cells and its preparation method and application, especially it is related to the lineage-specific differentiation of pluripotent or multipotent stem cells, specifically related to a kind of RIC cell obtained from stem cell differentiation and its preparation method and application.