497results 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 invention discloses a vascularized tissue culture chip as well as a construction method and application thereof, and belongs to the technical field of biological tissue engineering and biological medicines. The vascularization tissue culture chip is formed by stacking five layers of sub-chips with different function requirements, a fluid channel and a culture chamber are formed, a highly integrated system is formed, and the design of a micro-column array further optimizes the fluid dynamic characteristics and reduces the dead angle effect of fluid. In-vivo microcirculation can be simulated, and a stable and controllable nutrition and metabolite exchange environment is provided for skin and liver tissue. The vascularized tissue culture chip provided by the invention provides a powerful research platform for a plurality of biomedical research fields such as personalized medical treatment, disease modeling, drug screening, toxicity testing, disease model research and the like.

The invention relates to human hepatocyte organoids. In particular, hepatocyte organoids with modified genes involved in lipid homeostasis. The invention also relates to the use of such organoids for modelling diseases involving steatosis. Also included is the use of such organoids as models of fatty liver, such as caused by genetic as well as diet related fatty liver disorders and their use in discovery of novel drugs for treating fatty liver and related disorders. The invention further relates to agents for reducing steatosis in subjects having similar modifications as the hepatocyte organoids of the invention. The invention also relates to methods of reducing or preventing steatosis, treating or preventing cardiovascular disease; treating or preventing NAFLD, and preventing and / or reducing the risk of NASH and / or cirrhosis by increasing FADS2 activity in a subject. Also included is a FADS2 agonist, such as a small molecule, nucleic acid or polypeptide for use in said methods.

The invention belongs to the technical field of biomedical engineering and microfluidics, and particularly discloses a vascularized organ chip and method for large-scale culture, the vascularized organ chip comprises an upper runner layer and a lower culture layer, and the runner layer and the culture layer are fixedly sealed; a liquid storage pipe matched with the flow channel layer is adhered to the upper surface of the flow channel layer; according to the vascularized organ chip, the flow channel layer is used for pouring a culture solution to provide fluid stimulation and nutrients for cells; the three parallel-connected large-scale culture chambers of the culture layer are used for culturing cells, so that blood circulation of a microvascular system in a human body can be simulated, and a microenvironment in tissues can be reproduced more truly; meanwhile, the requirement for culturing a large number of tissue cells through the chip can be met, and the cell yield and the experiment scale can be effectively increased.

The present invention relates to an organ-on-a-chip with an inverted microweir structure as a cell co-culture platform and a use thereof. By using the multi-organs-on-chip provided by the present invention, it is possible to derive more accurate efficacy and toxicity results for cells according to drug treatment. In particular, when a drug is converted into a metabolite through liver metabolism in a liver-heart co-culture system, the resulting cardiotoxicity and drug responses across multiple organs can be predicted, and thus the chip can serve as an alternative to animal testing, significantly reducing the cost and time required for new drug development and drug screening. Additionally, the chip can be advantageously utilized in studies of cellular microenvironments and research on other organ-on-a-chip systems.

Provided are a liver precursor cell and a construction method therefor and the use thereof. The method for constructing liver precursor cells comprises the steps of: S0, providing liver precursor cells, a mixed solution and a reprogramming culture medium, wherein the mixed solution comprises a serum-free culture medium, polyethyleneimine and a CLP plasmid; S1, carrying out a transfection treatment on the liver precursor cells and the mixed solution to obtain transfected liver precursor cells; and S2, placing the transfected liver precursor cells into the reprogramming culture medium for subculture to obtain immortalized liver precursor cells. The method solves the problems that in the prior art, liver precursor cells cannot be transformed into immortalized cell strains by means of immortalizing genes and cannot be continuously passaged in vitro.

The invention discloses a liver progenitor organoid culture medium and a culture method, and belongs to the technical field of organoid culture. The culture medium comprises a basic culture medium, Jagged-1, Dexamehysone, TGF (Transforming Growth Factor)-alpha, beta-mercaptoethanol, BMP4 (Bone Morphogenetic Protein 4), FGF2 (Fibroblast Growth Factor 2), specific addition factors Glutamax, HEPES (Hydroxyethyl PES), Nicotinamide, Penicillin and the like. The liver progenitor organoid is constructed by using the liver tissue for the first time, continuous growth and efficient proliferation of human liver progenitor cells in vitro can be promoted under the condition that gene editing is not carried out, meanwhile, the pluripotency of the liver progenitor organoid is maintained, and the consistency of specific marker expression is maintained; and a novel batch model is provided for the evaluation of hepatotoxicity at the present stage. From the long-term view, a foundation is laid for constructing a liver organ model with more advanced various differentiated cells.

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 method for realizing in-vitro amplification of adherent cells by utilizing a stirring type bioreactor matched with a three-dimensional porous hydrogel carrier. The method comprises the following steps: sterilizing a hydrogel carrier and a stirred bioreactor, adding a cell culture medium, ventilating, inoculating cells, starting the stirred bioreactor for culturing, observing the color change of a culture solution in the culture process, and changing the solution when the culture solution turns yellow, thereby ensuring the nutrition required by cell culture. The hydrogel with high biocompatibility is used as a carrier, a porous three-dimensional growth environment is provided, cells can be attached to the interior of the hydrogel carrier, the cell growth space is sufficient, the cells can be cultured for more generations, the number and density are greatly increased, the amplification multiple can reach 10 times or above, the production efficiency is improved, and the production cost is reduced.

Disclosed are compositions, in particular, organoid compositions, derived from more than one donor cell. Further disclosed are methods of making compositions, for example, organoid compositions, that comprise a differentiated cell population derived from more than one donor cell. Donor cells may include, for example, a precursor cell such as an embryonic stem cell or other precursor cell. The disclosed methods use synchronization conditions to produce a synchronized pooled-precursor cell population, which may then be differentiated into an organoid composition. Methods of using the compositions are also disclosed.

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.

The invention discloses a preparation method and application of a cell-loaded porous 3D scaffold. The preparation method comprises the following steps: S1, preparing printing ink; s2, the printing ink prepared in the step S1 is added into 3D printing equipment, and a 3D support is printed according to a designed model; the printing ink comprises a main material, an auxiliary material and a cell-loaded suspension, the carrier is applied to dynamic and static culture, testing or screening of carrier cells. The specific surface area of the 3D scaffold is increased to the maximum extent, the dynamic and static culture, test and screening functions of the carrier cells are realized, and the scientific research efficiency is improved.

The present invention relates to a specific liver microtissue comprising at least three different phenotypes of liver cells obtained from induced pluripotent stem cells encapsulated in a single three-dimensional closed microcompartment. The invention also relates to a method for preparing the liver micro-tissue and application of the liver micro-tissue in treating or preventing liver failure.

The invention provides a method for preparing a DC vaccine based on exosomes from liver cancer organoid sources. The method comprises the following steps: S1, constructing a liver cancer organoid model; s2, extracting an exosome from the liver cancer organ model; s3, culturing DC cells of the liver cancer patient; s4, co-culturing the exosome obtained in the step S2 and the DC cells cultured in the step S3, and then harvesting mature DC cells. Exosome components containing liver cancer tumor antigen information are extracted and then added into a DC cell culture medium for antigen loading, and finally mixed immune cells activated by DC cells are added into autologous liver cancer organs of a patient, so that specific killing of the liver cancer organs can be realized.

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.

[Solution] The production and maintenance of hematopoietic stem cells in in vitro culture systems has proven difficult to achieve. Disclosed herein is a hematopoietic stem cell composition derived from organoids, such as liver organoids. These organoids also contain rare types of immune cells that have not yet been fully characterized due to the difficulty of isolating said immune cells from biological samples. Disclosed herein is a method for producing said hematopoietic stem cells and immune cells from organoids, as well as a method for expanding hematopoietic stem cells from other sources using these organoids.

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.