4101results about "3D culture" patented technology

The invention provides a microfluidic chip, system and method for osteochondral organs, and relates to the technical field of organoid microfluidic chips, the chip comprises a bone microvascular endothelial cell channel, a bone compartment, a cartilage compartment and a synovial fluid channel which are arranged in parallel and in sequence, and permeation areas between the channel and the compartments and between the compartments. According to the scheme, the complex structure and function of a human osteochondral system are effectively simulated, the physiological and pathological processes of human bone joints can be simulated more truly in an in-vitro environment, and a method for establishing a highly-simulated femoral head necrosis model and osteoarthritis model based on the chip is provided. The problems that a traditional model is insufficient in physiological environment simulation, lack of control over dynamic culture conditions, tedious and time-consuming in the construction process, difficult to support high-throughput experiments and the like are solved.

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 compositions (e.g., pharmaceutical compositions), uses, methods, non-therapeutic and therapeutic methods related to senolytic agents encapsulated or comprised in Muse cell derived extracellular vesicles (e.g. exosomes). Such compositions and methods are useful for specifically killing senescent cells and reverting any symptoms, disorders or diseases associated with senescence.

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 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 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 relates to a cell culture microcarrier as well as a preparation method and application thereof. The cell culture microcarrier comprises a polymer porous microsphere with an inside-outside through pore channel structure. The cell culture micro-carrier comprises a biodegradable micro-carrier and also comprises a non-degradable micro-carrier, so that the growth of cells in the carrier is facilitated, and the mass transfer of nutrient substances and metabolites is also facilitated, thereby promoting the growth of the cells, enabling the cultured cells to keep good activity and function, and being high in proliferation efficiency; the method is suitable for three-dimensional culture of somatic cells, stem cells, tumor cells and the like.

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

The invention discloses hyaluronic acid / alginic acid dual-network hydrogel as well as a preparation method and application thereof. The hyaluronic acid / alginic acid double-network hydrogel comprises a first network and a second network which are not crosslinked with each other, and the first network and the second network are distributed in the hydrogel in an interpenetrating manner; the first network comprises an ionic cross-linked network of alginic acid with cell adhesion sites and calcium ions, and the second network comprises a chemical cross-linked network formed by covalent cross-linking of methacrylic anhydride modified hyaluronic acid. The hyaluronic acid / alginic acid double-network hydrogel disclosed by the invention is short in curing time, supports three-dimensional spreading of cells and formation of capillary lumens, has good mechanical stability, and has good application prospects in the aspects of three-dimensional culture of the cells, tissue engineering construction and the like.

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 bone organ model as well as a construction method and application thereof, and the method comprises the following steps: arranging a biological material with a micropore array in a cell culture plate to obtain a bone organ chip; suspending the initial stem cells in a culture medium, and promoting the cells to form a spherical structure through cell aggregation or by using a non-adhesive culture surface, so as to obtain cell spheres; and culturing the cell spheres in the bone organ chip by adopting the culture medium for constructing the bone organ model to obtain the bone organ model. The skeletal organ is successfully constructed through in-vitro self-organized 3D co-culture initial stem cells including embryonic stem cells, induced pluripotent stem cells, bone marrow mesenchymal stem cells, osteoblasts, bone cells, all levels of osteogenic precursor cells, osteogenic-like cells and MC3T3-E1 cells, and the skeletal organ can become an important tool for researching and treating bone related diseases.

The invention relates to a method for induced differentiation of skeletal muscle cells by porcine embryonic stem cells. Comprising the following steps: (1) constructing a PAX7-overexpressed porcine embryonic stem cell stable cell line; (2) induced differentiation of skeletal muscle cells; (3) performing immunofluorescence identification; wherein the step (1) specifically comprises the following steps: 1.1, constructing a DOX (deoxyribonucleic acid) induced PAX7 expression plasmid PiggyBac-TRE3G-EF1a-PAX7; 1.2, performing electrical transformation on the porcine embryonic stem cells cultured for two days, and adding Buffer E into an electrical transformation cup; taking Buffer R, mixing plasmid and PBase, uniformly mixing, and re-suspending cell precipitate; 1.3, after electrotransformation is finished, the cells are planted on the feeding layer, Y-27632 is added into the culture medium, and the culture medium without Y-27632 is replaced with the culture medium without Y-27632 on the second day; and 1.4, after culturing for 3-4 days, selecting the porcine embryonic stem cells which are successfully subjected to electrotransfection for amplification culture, and naming PAX7-PESCs.

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.

A method for prepring a porous aerogel scaffold includes: adding a photoinitiator and polyethylene glycol diacrylate in a buffer solution, dissolving by heating and evenly mixing, adding Pluronic F127 into the mixed solution, and standing at a low temperature to obtain an aerogel scaffold material; printing a hydrogel scaffold by using a 3D printing technology, and performing UV irradiation so that a cross-linking of the hydrogel scaffold is caused to form a three-dimensional scaffold with a stable structure, performing low-temperature soaking to remove Pluronic F127, and then freeze drying the three-dimensional scaffold to obtain the porous aerogel scaffold. Wherein, Pluronic F127 serves as a sacrificial material which is removed after the 3D printing of the hydrogel scaffold is completed, and then a porous structure can be formed in the scaffold in combination with a freeze drying technology, which facilitates the survival, growth and proliferation of cells during the three-dimensional culture.

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 present invention relates to an intestinal epithelium model accurately reproducing the pathophysiological mechanisms observed in vivo in the context an inflammatory condition of the intestinal barrier. The model according to the present invention comprises two compartments separated by a semi-permeable membrane. The first compartment, corresponding to the apical pole of the intestinal epithelium, comprises a coculture of Caco-2 cells differentiated into enterocytes and HT29-MTX cells differentiated into goblet cells. The second compartment, corresponding to the basolateral pole of the intestinal epithelium, comprises a culture of THP-1 monocytic cells differentiated into macrophages. The model according to the present invention is characterized in that the cells contained in the first compartment produce interleukin 6 (IL-6) at a concentration of more than 100 pg / ml and interleukin 8 (IL-8) at a concentration of more than 150 pg / ml, and the cells contained in the second compartment produce tumour necrosis factor α (TNF-α) at a concentration of more than 40 pg / ml and interleukin 1β (IL-1β) at a concentration of more than 90 pg / ml. The present invention also relates to the method for obtaining this model and also to a method for selecting candidate compounds for the treatment of inflammatory bowel diseases by verifying their ability to stop the inflammation as soon as the permeability is increased, or verifying the restoration of the permeability and of the mucus layer.

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 discloses a construction method and application of a mouse airway organoid inflammation model, and relates to the technical field of biology. The construction method comprises the following steps: (1) taking a mouse airway tissue, digesting with digestive juice, and centrifuging to obtain a cell suspension; (2) centrifuging the cell suspension, discarding the supernatant, adding matrigel, uniformly mixing, incubating, and culturing by using an airway organoid complete medium; (3) adding a cell recovery solution into the cells cultured in the step (2), incubating, degumming, centrifuging, discarding supernatant, adding matrigel, uniformly mixing, incubating, and adding an airway organ complete culture medium for continuous culture; and (4) adding 1-20 [mu] g / ml of LPS into the airway organoid complete culture medium, and incubating. The AO constructed by the invention covers four main cell types of cilia cells, goblet cells, basal cells and rod-like cells, causes epithelial injury in the AO and expression of AO inflammatory factors, and can provide reference for development of CRD new drugs.

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.

In various aspects, the present invention provides a composition having orthogonally tunable elasticity and viscosity. In some embodiments, the composition comprises an interpenetrating polymer network comprising viscous and elastic polymer networks that are not covalently bonded. In some embodiments, the composition is a viscoelastic hydrogel. In some embodiments, the composition mimics the viscoelasticity of at least one organ, organoid, or tissue of interest (e.g., brain, pancreas, brain tumor, etc.). In various aspects, the present invention provides a device (e.g., 3-dimensional cell culture platform) comprising said composition. In some aspects, the present invention provides a method of preparing said devices. In some embodiments, the method comprises bioprinting.

Methods and systems for performing a bioproduction process are described, the bioproduction process comprising the production of a transformed cell population. The methods include the steps of culturing a cell population at least partially embedded in a hydrogel matrix forming a three-dimensional structure, the hydrogel matrix also comprising a transforming agent.

A microfluidic device, a microfluidic system comprising said microfluidic device, and to a method of culturing biological material.

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.