264results about "Hepatocytes" patented technology

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 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 application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

Techniques for generating and utilizing adult hepatocyte organoids are described. An example method includes generating an organoid by culturing hepatocytes obtained from an adult donor in a hepatocyte culture medium. The hepatocyte culture medium includes a basal medium and 5 μM of a TGF-β inhibitor.

The invention relates to the technical field of polypeptides, in particular to an enzyme-mimetic polypeptide, hydrogel, a preparation method and application, the enzyme-mimetic polypeptide selects a peptide fragment I and a peptide fragment II with biocompatibility and degradability as basic sequences, and the peptide fragment I and the peptide fragment II form a beta folding unit, so that the stability of a polypeptide structure can be enhanced; then, the peptide chain is driven to be self-assembled to form a regular nanofiber structure; according to the invention, the characteristic that the biomimetic polypeptide and metal ions are subjected to a complex reaction to form a chelate is utilized, the transparent and stable hydrogel can be rapidly formed without adding a gelatinizing agent, and TEM, SEM and rheological characterization are carried out on the hydrogel to find that the hydrogel containing the biomimetic polypeptide has a porous nanofiber network structure, so that the biomimetic polypeptide can be used for preparing the hydrogel. Meanwhile, the biomimetic polypeptide hydrogel has the same flexibility as natural tissues, and the characteristics indicate that the biomimetic polypeptide hydrogel is expected to have wide application prospects in the aspects of biomimetic materials, antibiosis, cell culture, drug carriers and the like.

The invention discloses a perfusion-free parallel double-cell intestine-liver chip system and application thereof in drug uptake evaluation, and belongs to the field of biomedical engineering and organ chips. The chip disclosed by the invention is simple in structure, free of an external pump and a complex pipeline, capable of realizing fluid circulation through swinging, convenient to operate, low in cost and small in pollution risk, and also capable of promoting cell differentiation and optimizing hepatocyte functions. The chip integrates a parallel intestinal cell and a liver cell which are communicated through a shared channel, can continuously simulate intestinal absorption and liver first-pass metabolism, and is closer to an in-vivo physiological state. The device adopts a detachable transparent modular design, can be repeatedly used, supports multi-group parallel operation, can improve experimental flux and result repeatability, and is suitable for in-vitro verification of drug absorption, permeability and metabolism evaluation.

The invention discloses a construction and evaluation method of a liver injury organoid model, which comprises the following steps: (1) selecting mouse liver cells, perfusing a mouse with pancreatin through a sterile operation technology, removing blood in the liver, collecting liver tissues, and carrying out fine enzymolysis digestion treatment to obtain a single-cell suspension; (2) inoculating the single-cell suspension into a three-dimensional culture system, and carrying out preliminary stimulation by adopting a culture medium combined with growth factors and inhibition small molecules, so as to promote the growth, proliferation and differentiation of cells in a three-dimensional space; according to the cell growth condition, subculture operation is carried out in good time, so that continuous culture and amplification of liver organs are ensured; and (3) exposing the maturely cultured liver organoid in a stimulating factor for simulating a liver injury pathological environment, and constructing different types of liver injury organoid models. According to the liver injury organ-like model construction method, model construction can be completed in a short time, the research period is greatly shortened, the efficiency is high, and the cost is low.

Disclosed herein are three-dimensional culture system for producing cartilage and / or selecting therapeutic agents for promoting cartilage regeneration.

The present disclosure provides methods of making tissue-derived epithelial organoids. In particular, the present disclosure provides a tissue-derived epithelial organoid embedded in a hydrogel suspended in a culture medium. The present disclosure further provides methods of using such organoids.

Systems and methods consistent with the present invention generally relate to microorganospheres (MOSs), and methods and apparatuses for forming and using MOSs. More particularly, in some embodiments, systems and methods consistent with the invention relate to the methods and apparatuses for forming and using MOSs generated from hepatocytes. MOSs that are generated from hepatocytes are suitable for testing liver toxicity and drug induced liver injury effects of various agents.

The invention relates to a cellulose-based hydrogel prepared based on a cellulose copolymer, a cellulose derivative copolymer or a blend thereof, ultramicro fibers of cellulose form a net structure, and the cellulose-based hydrogel has a highly-ordered three-dimensional micro-nano structure, is better in transparency and is more beneficial to observation of a cell state. The cellulose-based hydrogel is suitable for culture of 3D cell culture tissues and organoids, the stem cells cultured by the cellulose-based hydrogel can form cluster culture stem cells and organoids, the operation is simple, compact fused monolayer endoderm cells are formed, non-directional differentiation of the stem cells can be inhibited, and the formed organoids are thick in vesicle wall and are in a solid shape. And the obtained product is a differentiated mature organ and can be used for high-throughput drug screening, drug toxicity testing and regenerative medicine.

Featured here are methods for improving tissue or organ preservation and utilization through mitochondrial transplantation.

The present invention relates to specific liver microtissues comprising hepatocytes of at least three different phenotypes derived from induced pluripotent stem cells encapsulated in a single three-dimensional closed microcompartment. The present invention also relates to methods for preparing such liver microtissues and their use in the treatment or prevention of liver failure.

Production and maintenance of hematopoietic stem cells in in vitro culture systems has proven to be elusive. Disclosed herein are hematopoietic stem cell compositions that originate from organoids such as liver organoids. These organoids also comprise a rare type of immune cell that is not yet fully elucidated due to the difficulty in isolating said immune cell from biological samples. Also disclosed herein are methods of producing said hematopoietic stem cells and immune cells from organoids, as well as methods of expanding hematopoietic stem cells from other sources using these organoids.

The invention relates to the field of cell culture, in particular to a liver organ model culture device and method.The liver organ model culture device comprises a temperature control box and a box cover arranged on the temperature control box, a pressing column is slidably installed in the middle of the box cover in a penetrating mode, a column spring is wound around the outer side of the pressing column, and one end of the column spring is connected with the upper end of the pressing column; the other end of the column spring is fixed to the upper end of the box cover, a culture piece is arranged at the position, located on the inner side of the box cover, of the upper end of the temperature control box, a plurality of liquid changing holes are formed in the upper end of the box cover in a penetrating mode, a bearing frame is placed at the upper end of the box cover, and a plurality of liquid injection pipes are embedded in the bearing frame in a penetrating mode; a plurality of liquid suction pipes are embedded in the bearing frame in a penetrating mode, and the liquid injection pipes are located on the sides of the liquid suction pipes. According to the invention, the culture effect is improved, the liquid changing efficiency is also improved, and meanwhile, the gel matrix block is prevented from being damaged during liquid changing.

The invention belongs to the technical field of bioengineering, particularly relates to the field of tumor animal model research, and particularly provides a primary in-situ hepatocellular carcinoma animal model and a construction method thereof, and the primary in-situ hepatocellular carcinoma animal model is characterized in that primary in-situ hepatocellular carcinoma is formed in an animal liver; the primary in-situ hepatocellular carcinoma is formed in a primary in-situ manner by introducing a gene-edited liver organ into an animal liver tissue. The innovative hepatocellular carcinoma model construction technology promotes the primary in-situ development of long liver organs after gene editing into hepatocellular carcinoma, not only is the preparation period shorter, but also the tumor formation efficiency is higher, the occurrence and development, pathological conditions and clinical drug response of liver cancer patients can be effectively simulated, the full simulation of the tumor microenvironment is realized, and the application prospect is wide. And the development and research of new drugs and new treatment methods can be better guided.

A novel method of inducing or producing hepatocytes from human induced pluripotent stem cells at an unprecedented efficiency and functionality. The core of the invention is the use of experimentally discovered mRNAs at multiple critical differentiation decision points along a pluripotent to mesendoderm to endoderm to hepatocytes pathway in a previously unknown manner.

The present disclosure provides porcine thymolivers, methods of making thereof, and methods of using thereof. The present disclosure further provides methods of treating a disease (e.g., a liver disease) in a human subject in need thereof comprising transplanting of a porcine thymoliver.

The application provides a preparation method of organoids based on a single cell chip, which comprises microfluidic chip preparation, a two-water-phase solution preparation, a single cell suspension preparation, microfluidic manipulation, single cell-loaded microgel generation and organoid construction and the like. The application takes a microfluidic chip integrated with a normally closed pneumatic pump valve as a technical platform, takes a single cell suspension pre-incubated with a crosslinking agent as a chemical reaction core, takes a water-in-water droplet as a forming template, and realizes efficient in-situ loading of single dispersed seed cells in a micro hydrogel carrier through a one-step method, and simply removes empty droplets without cells in a subsequent transfer step. The single cells in the microgel can be cultured, expanded and self-organized to form required organoids. The application improves the consistency of the starting point of the organoids, has the high-throughput characteristics of the droplet microfluidic technology, realizes efficient and highly reproducible in-vitro construction of the organoids, and can play a great role in the fields of basic research and transformation application related to the organoids.

The present invention relates to methods for the in vitro production of hepatocellular organoids from primary human hepatocytes (PHHs). The invention also relates to a cell culture system for the in vitro expansion of functional fetal, neonatal, pediatric and adult PHHs and the mass production of organoids, and to the use of said cell culture system for the in vitro mass production and expansion of functional adult PHHs and / or the formation of hepatocellular organoids.

To provide a method for producing a cell aggregate that can be used as a liver model in a state more similar to that in vivo.SOLUTION: The present invention relates to a method for producing a cell aggregate having a bile canaliculus, the method including: a contact step for bringing heparin and cells, including hepatocytes, into contact with each other in an aqueous medium; and a culturing step for culturing the cells that have been brought into contact with heparin to form a cell aggregate.SELECTED DRAWING: None

The present invention relates to methods for the in vitro production of a hepatocyte organoid from primary human hepatocytes (PHH). The present invention further relates to a cell culture system for the in vitro expansion of functional foetal, neonatal, paediatric and adult PHH and the mass generation of organoids and use of the cell culture system for the in vitro mass generation and expansion of functional adult PHH and / or formation of hepatocyte organoid.

A method for inducing conversion from non-hepatic stem cells or non-hepatic progenitor cells into hepatic stem cells or hepatic progenitor cells, which comprises introducing any of the following combinations into the non-hepatic stem cells or non-hepatic progenitor cells: (a) a combination of HNF1, HNF6 and FOXA; (b) a combination of HNF1 gene, HNF6 gene and FOXA gene; (c) a combination of HNF1, MYC and FOXA; or (d) a combination of HNF1 gene, MYC gene and FOXA gene.

This invention belongs to the field of hydrogel materials and relates to a method for preparing a hydrogel, comprising the following steps: dissolving bisphosphonic acid-modified pectin in water or an aqueous solution, adding a divalent metal cation solution to mix and obtain a precursor solution, adding a gelling solution to initiate cross-linking and form a hydrogel, wherein the gelling solution is a solution containing monovalent metal ions with a pH of 7-9. The hydrogel has broad application prospects in the field of ready-to-use reagents. Adding the precursor solution to a cell culture device and lyophilizing it can be used to prepare ready-to-use reagent kits. Dropping a cell suspension into the ready-to-use hydrogel reagent kit and adding cell culture medium yields a hydrogel encapsulated with cells for three-dimensional culture.

The invention provides an organoid culture medium, an organoid culture method and application of the organoid culture medium, the organoid culture medium comprises a first-stage culture medium, the first-stage culture medium comprises a first-stage basic culture medium and first-stage addition factors, and the first-stage addition factors comprise N-acetylcysteine, nicotinamide, a ROCK inhibitor, Noggin, R-spondin, EGF, HGF, Gastrin, FGF10, Blebbistatin, Forskolin, A83-01, LDN193189, DAPT, TGF-alpha and UDAC. By using the organoid culture medium, the liver organoid and the fatty liver organoid can be effectively obtained, and the obtained fatty liver organoid has obvious fat deposition, is sensitive to fatty liver treatment drugs, highly restores the physiological status when human body fatty liver diseases occur, and has high application value.

The invention belongs to the field of biomedicine, and particularly relates to a construction method of an in-situ biological 3D printing artificial liver patch for treating hepatic failure. The main component of the artificial liver patch disclosed by the invention is composite hydrogel, and the composite hydrogel comprises hyaluronic acid (HA), gelatin (Gelatin), sodium alginate (sodium alginate) and methacrylated gelatin (GelMA). The composite hydrogel has good 3D printing performance, large-scale construction of the artificial liver patch can be achieved through biological 3D printing, and a potential solution is provided for transplantable biological artificial liver tissue. The artificial liver patch adopts an extrusion type printing mode, the forming speed is high, internal liver cells can still keep high activity, reconstruction of liver basic functions (glycogen storage, drug metabolism, secretory protein synthesis and the like) can be guaranteed, and the artificial liver patch has important potential in treatment of liver failure diseases.

Disclosed herein are improved methods of expanding cell populations and methods of producing organoids from pluripotent stem cells, as well as compositions and uses thereof, compositions comprising liquid components that can be used to carry out these methods for supplementing amino acids, and kits comprising components for carrying out these methods and / or products of using these methods. These methods can be carried out without the need for xenogeneic and ambiguous basement membrane matrices for cell culture, which enables the achievement of good production specification (GMP) compliant methods for producing organoids for various uses such as clinical screening and treatment, methods for treating liver-related diseases or conditions and methods for screening candidate compounds or compositions for treating liver-related diseases or conditions are included.

The present invention provides means for producing an organoid close to an organ in a living body and capable of secretion of a plasma protein and immune response. A matrix composition of the present invention provided as such means includes: (1) a first matrix containing one or more cells selected from the group consisting of vascular cells, nerve cells, and blood cells; and (2) a second matrix containing to cells constituting an organ and / or an organoid, in which the first matrix envelops the second matrix, and the first matrix has at least one opening.

The invention relates to the technical field of cell culture, in particular to nano liver stem cell peptide as well as a preparation method and application thereof. The preparation method of the nano liver stem cell peptide comprises the following steps: (1) mixing induced pluripotent stem cells with a first induced differentiation culture medium, and culturing for 3-5 days to obtain an intermediate cell 1; (2) inoculating the intermediate cell 1 into a second induced differentiation culture medium, and culturing for 3-5 days to obtain an intermediate cell 2; and (3) inoculating the intermediate cell 2 into a third induced differentiation culture medium, and culturing for 3-5 days to obtain the liver stem cell, and (4) after the liver stem cells are subjected to ultrafiltration, taking substances of which the molecular weight cutoff is less than or equal to 50kDa, centrifuging, taking precipitates, and drying to obtain the liver stem cell peptide. The liver stem cell peptide prepared by the method has the particle size of 1-20nm, can target the liver, has a good effect in treatment of liver injury, and does not generate drug resistance.

A method for producing a metabolism-activated liver organ, comprising a step for culturing primary hepatocytes or hepatocyte-like cells in a differential medium to obtain a metabolism-activated liver organ, the differential medium is a medium containing a glucocorticoid receptor agonist and one or two selected from the group consisting of a growth hormone receptor agonist and a prolactin receptor agonist.