2870results about "3D culture" patented technology

A system for growing cells comprising a bioreactor chamber for growing the cells, a delivery system delivering a perfusion solution to the bioreactor chamber for perfusion of the perfusion solution through the cells, a dialysis system having a dialyzer, a dialysate for performing a dialysis and a filter for reducing ammonia content in said dialysate, and a controller that circulates the perfusion solution through the dialyzer and the dialysate through the filter.

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 present disclosure provides systems and methods for processing cells. The systems of the present disclosure may include a compartment, a three-dimensional (3D) matrix, a fluid flow path, an analysis unit, a preservation unit, or any combination thereof. The methods of the present disclosure may process cells via, for example, expansion, culturing, gene delivery, activation, or any combination thereof. The systems and methods described herein may be useful for producing cell products including, for example, cell therapy products.

The invention belongs to the technical field of tissue engineering, and particularly discloses a preparation method of a 3D full-thickness skin model. The preparation method provided by the invention comprises the following steps: constructing a corium layer, constructing an inner epidermal layer through first epidermal inoculation, constructing an epidermal layer through second epidermal inoculation and obtaining a complete 3D full-thickness skin model through co-culture. The 3D full-thickness skin model obtained through the preparation method is distinct in gradation, uniform in thickness, free of shrinkage and stable in quality, the epidermal layer and the corium layer are uniformly fused, and the 3D full-thickness skin model is suitable for skin function research and sample evaluation. Meanwhile, the preparation method provided by the invention is simple in process and convenient to operate, does not need additional reagents or anti-shrinkage tools, can meet the requirements of large-scale industrial production, and has good production feasibility and market application prospects.

The invention provides human placenta-derived vasogenic stem cells (hPASCs) and application thereof. The human placenta-derived angiogenic stem cells (hPASCs) disclosed by the invention are preserved in China Center for Type Culture Collection, Wuhan University, Wuhan, China; the preservation date is April 2, 2025; the preservation number of the strain is CCTCC (China Center For Type Culture Collection) NO: The hPASCs disclosed by the invention contain angiogenic cell subgroups, have stem cell stemness, have an angiogenesis effect in vivo and in vitro, and can be used for treatment of diseases such as cardiovascular diseases and vascularization construction of artificial organs and tissues.

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 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.

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.

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

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

The invention relates to 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 discloses zwitter-ion supramolecular hydrogel based on anion-pi as well as a preparation method and application of the zwitter-ion supramolecular hydrogel. The hydrogel is formed by self-assembly of a zwitterionic copolymer and a p-styrenesulfonic acid copolymer. According to the supramolecular hydrogel disclosed by the invention, the supramolecular hydrogel can be rapidly formed only by simply mechanically mixing the zwitterionic polymer copolymer and the p-styrene sulfonate copolymer solution, and the supramolecular hydrogel is endowed with excellent injectability and self-repairing performance due to the dynamic cross-linking characteristic. The hydrogel is composed of zwitterions with excellent immunocompatibility and poly (p-styrene sulfonate) with heparin-imitating performance, so that the hydrogel has excellent immunocompatibility, can play a role in various in-vivo application scenes, and in addition, the hydrogel can be used for preparing an anti-inflammatory drug. The dynamic cross-linked network based on anion-pi interaction and the heparin-like structure are very beneficial to protein preservation, and have wide application prospects in the fields of protein delivery and cell encapsulation.

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.

A robust nasal organoid culture system and fully characterized 2D and 3D in vitro produced organoids are provided, as well as methods of using the organoids. The nasal organoids comprise a population of differentiated nasal epithelial cells having cellular heterogeneity and molecular markers similar to mature nasal epithelial tissue. Improved methods for producing two-dimensional and three-dimensional nasal organoids are also provided. These methods generally require a two-phase culture system, i.e., amplification and differentiation culture. The provided nasal organoids are suitable for reproducing nasal tissue complexity, for mimicking respiratory diseases, studying barrier function and drug delivery, high throughput drug screening, and assessing the effects of pathogens and environmental toxins on nasal respiratory epithelium.

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

The invention relates to a triple-hole paper-based three-dimensional cell culture chip as well as a preparation method and application thereof. The chip comprises a paraffin hydrophobic region and a paper-based hydrophilic region. The preparation method mainly comprises the following steps: 1) designing a mask plate; 2) cutting the mask plate by laser; and 3) preparing the triple-hole paper-based chip by hot pressing. The thickness of the chip is 100-200 [mu] m, the porous fiber structure of the paper-based material allows transportation of oxygen and waste, and a three-dimensional microenvironment for in-vivo cell growth can be simulated. Meanwhile, a transparent and liquid-tight supporting layer is fused on the lower layer of the paper-based chip, so that leakage of cells inoculated on the upper layer of the paper-based chip is avoided. The paper-based chip is embedded into a 6-pore plate, and bladder cancer cells (5637), mouse astrocytoma (U87) and human breast cancer cells (MCF-7) are inoculated in a paper-based hydrophilic region respectively. The cell compatibility of the paper-based chip and the growth behaviors of three different types of cells in three-dimensional paper fibers are investigated. It is proved that the chip can be used for researching three-dimensional construction and growth of human and mammalian cells, and the difference between three-dimensional culture and two-dimensional culture is revealed by detecting the proliferation capacity and survival rate of different cells.

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

The present invention relates to inflammatory bowel disease model production using patient-derived intestinal organoids, and use thereof, wherein the inflammatory bowel disease model produced by an optimized production method for inflammatory bowel disease model production can be used as a patient-customized bowel disease model by constructing an evaluation method for inflammatory bowel disease drugs.

A method for constructing a microfluidic bone organoid-on-chip can continuously observe the impacts of different external factors on a bone tissue structure based on the “ternary regulation theory” of bone angiogenesis-bone resorption-osteogenesis coupling. The method includes: (1) building a visual three-dimensional cell cultivation platform to continuously observe impacts of different external factors on a ternary regulation theory based on angiogenesis of osteoclast precursors, osteoblasts and osteoclasts, such that a user is allowed to directly observe interactions among various kinds of cells; (2) designing microfluidic channels in a chip to allow a reagent involved to present a concentration gradient distribution in the chip; and (3) adding three bioinks including osteoblasts, osteoclasts and vascular endothelial cells respectively, so as to reflect the three cell components individually, in pairs, and as a whole in the chip. An apparatus for constructing a microfluidic bone organoid-on-chip is further provided.

The invention relates to a blood flow connection type multi-organ chip capable of being freely assembled and reused, the blood flow connection type multi-organ chip comprises a chip cover body, organ culture chambers, a chip matrix and a pressure-sensitive film, the chip matrix comprises a plurality of groups of organ culture units which are mutually independent, each group of organ culture units comprises at least two culture flow channels and a plurality of organ culture holes, and the culture flow channels are communicated with the pressure-sensitive film. The organ culture holes are sequentially distributed at intervals along the culture flow channel, the bottom end of each organ culture hole is communicated with the culture flow channel, each organ culture hole is used for accommodating an organ culture chamber, the culture flow channel is located at the bottom of the chip substrate, one end of the culture flow channel is communicated with the storage pool, and the other end of the culture flow channel is communicated with the connecting port; the chip cover body is arranged at the top of the chip base body and completely covers the storage pools and the organ culture holes, the pressure-sensitive film covers the bottom of the chip base body to seal the culture flow channels, and in each group of organ culture units, a circulation path is formed between at least two culture flow channels through the storage pools. The biological function of the human body is better simulated, and disease modeling and treatment strategy research are carried out.

The present invention provides a method and system for fabricating complex human tissues by integrating bio-electrospraying, cell electrospinning merged with 3D multi material microfluidic bioprinting, and future additions of magnetic and acoustic levitation technologies. The fabricated tissues serve as an alternative to animal testing in drug discovery processes, aiding in the prediction of human physiological responses to various drug compounds.

The invention relates to the technical field of biological products, and discloses a mesenchymal stem cell hair growth liquid and a preparation method thereof, and the mesenchymal stem cell hair growth liquid comprises the following components: hair follicle mesenchymal stem cells; a hair follicle mesenchymal stem cell exosome; growth factors: a vascular endothelial growth factor (VEGF), a fibroblast growth factor (FGFs), a keratinocyte growth factor (KGF-7) and interleukin-10 (IL-10); vitamins: vitamin E, vitamin B5, vitamin B2, vitamin B6, vitamin C and biotin; the plant extracts comprise a cacumen biotae extract, a polygonum multiflorum extract, a ginseng extract, a peppermint extract, a purslane extract and a rhodiola rosea extract; other active ingredients: tripeptide and digoxin; the auxiliary material is a hyaluronic acid-collagen peptide graft copolymer; and the balance of a phosphate buffer solution.

The invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized islet organoid and a construction method thereof. Aiming at the problems that the existing pancreatic islet organ vascularization mostly adopts a co-culture mode, the operation process is long, the difficulty is high, the randomness is realized, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized pancreatic islet organ. The method comprises the following steps of: sequentially inducing and culturing directional endoderm cells, original intestinal canal cells and posterior preintestinal cells, and adding a culture medium containing VEGF (vascular endothelial growth factor), BMP4 (bone morphogenetic protein 4) and FGF2 (fibroblast growth factor 2) for culturing in subsequent induced differentiation, so as to obtain the co-differentiated vascularized islet organ. Under the condition that exogenous endothelial cells are not added, the iPS cells are directionally differentiated to generate the vascularized islet organ containing beta, alpha and delta cells and vascular endothelial cells for the first time, the vascularized islet organ is more mature than non-vascularized islet organs and has a stronger insulin secretion function, and a new thought and an experimental basis are provided for realizing functional healing of diabetes mellitus.

The invention belongs to the technical field of biotechnology and animal models, and discloses a double-person-derived mouse model for simulating a tumor immune microenvironment and a construction method and application thereof. The method comprises the following steps: firstly, pretreating NSG immunodeficient mice by adopting low-dose whole-body irradiation in combination with double-antibody targeted bone marrow depletion, and transplanting CD34 + hematopoietic stem cells from the same human donor to complete human immune system reconstruction; separating tumor primary cells, tumor-related fibroblasts and tumor vascular endothelial cells of the same donor, performing three-dimensional co-culture to obtain homologous human tumor organs, and performing in-situ inoculation to immune reconstruction mice to obtain a target model. The core defects of MHC mismatching, low immune reconstruction efficiency, poor tumor immune microenvironment simulation degree, low clinical consistency and the like of an existing model are overcome, and the method can be used for tumor immune treatment drug screening, microenvironment mechanism research and personalized tumor treatment scheme verification.

The invention discloses a construction method of a homogenized self-assembled heart organoid, and belongs to the field of organoid. The preparation scheme of the heart organoid provided by the invention is controllable in method and simple in procedure, emphasizes the transition of differentiation from an iPSC 2D level to a 3D self-assembly heart organoid, overcomes the heterogeneity, including size, structure, function and even gene difference, of cross-batch or even same-batch organoid caused by an existing differentiation method, and has good cross-batch reproducibility. The constructed heart organoid contains various cell types and can continuously beat for at least 180 days in vitro, and the heart organoid in the same batch is uniform in height and stable in function. The preparation scheme does not need an engineering scaffold, matrigel and the like, emphasizes self-assembly of heart organs, and better accords with physiological characteristics of a human body.

Disclosed are methods of augmenting efficacy of oncology therapies by providing reprogrammed cells such as personalized regenerative cells that modulate the tumor microenvironment. In one embodiment, somatic cells are dedifferentiated into OCT-4 expressing cells and endowed with tumor homing properties through culture or gene engineering. Once a stable population of tumor homing cells is established, such cells are made to express immune stimulating agents constitutively, or selectively upon entering the cancer microenvironment. Selective expression of immune stimulatory agents can be induced by exposure to hypoxia, acidosis, immune suppressive signaling or inflammatory signaling.

The present disclosure relates to a method for producing an inner ear organoid with improved differentiation efficiency of hair cells. The inventors of the present disclosure have confirmed that when retinoic acid is treated at the stage of formation of the lamina propria during the production of an inner ear organoid, the differentiation potential of hair cells is excellent. Therefore, it is expected that the inner ear organoid produced according to the production method of the present disclosure can be advantageously utilized in hearing loss disease modeling, drug screening and regenerative medicine.

The invention belongs to the technical field of biological culture, and particularly relates to a three-dimensional dynamic biological culture device and method. The three-dimensional dynamic biological culture device comprises: a culture chamber unit, which comprises a porous membrane and a chamber body with a first opening, the porous membrane and the chamber body are detachably enclosed to form a first chamber, and the first chamber can accommodate a curable culture medium for providing a growth space for culturing cells; the flow path plate unit comprises a base part, the base part is provided with a circulation groove and a liquid storage opening matched with a liquid storage device storing a culture solution, and the liquid storage opening is communicated with the circulation groove; a plurality of mounting ports which can be matched with the culture room units are formed in the circulating groove and are used for communicating the first chamber with the circulating groove through the porous membrane; and the driving unit and the circulating groove form a circulating loop so as to drive the culture solution in the circulating groove to flow in the plurality of culture room units. According to the invention, dynamic culture of three-dimensional cells, cell spheres and organoid cells can be better realized.

The present invention provides: a drug which can improve the implantation rate of an embryo and can be used in conjunction with conventional drugs for infertility treatment; and a screening method and the like with which it is possible to reliably evaluate such a drug. Specifically, the integrin activator comprises: (1) a peptide composed of KFEERMRCKWMT, and (2) a peptide composed of KFEERMRCKWMT. (2) a peptide which is composed of KFEEERSRCKWMT; (3) a peptide comprising an amino acid sequence obtained by deleting, substituting and / or adding 1-3 amino acids in (1) or (2); or (4) a peptide obtained by binding any one of (1) to (3) to a membrane-permeable molecule. Provided are an artificial uterus and a screening method based on an implantation reproduction test system using a fertilized egg (for example, an embryo developed to an implanted blastocyst), the artificial uterus comprising: a hydrogel containing endometrial stromal cells and having an extracellular matrix as a main component; and a uterine epithelial organoid configured to be surrounded by the hydrogel.