1196results about "3D culture" patented technology

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

The invention provides a cryopreservation and resuscitation method for induced differentiation of pluripotent stem cells to metaphase cells and application, and the cryopreservation method comprises the following steps: (1) the initial cells are pluripotent stem cells, and are induced and differentiated to the metaphase; (2) recovering the cells by using a mild digestive enzyme, counting, and resuspending the cells in a cryopreservation solution; and (3) carrying out programmed cooling to-80 DEG C, and then transferring into liquid nitrogen for preservation for later use. According to the cryopreservation method and the cryopreservation liquid provided by the invention, the cryopreservation survival rate of the differentiated metaphase cells is obviously improved, apoptosis or irreversible stress injury induced by a traditional cryopreservation liquid is avoided, and the problems of large cell injury and low survival rate caused by using a general cryopreservation liquid in the prior art are obviously improved. The resuscitated cells keep good differentiation potential, can smoothly form a mature renal unit structure, and solves the problem of subsequent differentiation failure in the prior art.

Provided are chemically modified silk fibroin and a use thereof in cell and organoid culture. The chemically modified silk fibroin is a product obtained by sequentially carboxylating silk fibroin and modifying same by an organic amine having a phenol group. Also provided is a hydrogel based on the chemically modified silk fibroin. A hydrogel system has a hierarchical structure and mechanical properties similar to those of an extracellular matrix, and exhibits characteristics such as definite composition, controllable physicochemical properties, low cytotoxicity, good biocompatibility, and biodegradability, can support the growth and differentiation of cells and organoids, and is suitable as a matrigel for cell and organoid culture.

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

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

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

The present invention provides isolated human breast milk exosomes and compositions thereof. The invention also provides an in vitro method for producing the exosome, comprising generating lactating cells derived from human induced pluripotent stem cells (hiPSCs), expressing a human milk-like product from the lactating cells, and purifying the exosome from the human milk-like product.

The invention provides a method for evaluating drug enterotoxicity in vitro through an intestinal organ model, and relates to the field of biological medicine. Comprising the following steps: preparing six different culture media, culturing the hESC by using the different culture media, inducing the hESC to respectively perform directional endoderm cell differentiation, posterior intestinal bulb differentiation, intestinal organ differentiation and intestinal organ maturation, and evaluating the drug enterotoxicity by using a prepared intestinal organ model. The intestinal organ model prepared by the invention is similar to human intestinal cells in composition, can accurately reflect the toxic effect of the medicine in the physiological environment in the human body, can be repeatedly obtained in quantity, and avoids clinical risks and ethical constraints; the sensitivity of drugs with different enterotoxicity can be reflected through cell viability, and the method is suitable for rapid evaluation of in-vitro high-flux drug enterotoxicity.

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

The application discloses a construction method and application of a thymus organoid, and the method is used in combination with small molecule compounds and small molecule proteins to differentiate pluripotent stem cells into the thymus organoid under a suspension culture condition, and the method can efficiently obtain the thymus organoid which is closer to the natural thymus in structure and function.

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.

Disclosed are methods for producing controlled endometrial organoids using micropatterning techniques, and methods for using endometrial cells or organoids for diagnostic and predictive purposes, and for optimizing IVF and other infertility-related treatments, including embryo transfer and intrauterine insemination (IUI). These organoids grow in a spatially controlled manner and exhibit less heterogeneity than organoids grown directly in a three-dimensional culture. The specification also describes methods for predicting the success of embryo transfer or other IVF-related treatments, optimizing embryo transfer, improving embryo implantation in the endometrium, and performing embryo transfer in subjects. These methods include culturing endometrial organoids or endometrial epithelial cells, and contacting the endometrial organoids or cells with one or more blast-like cells. In some embodiments, the endometrial organoids or cells include endometrial cells obtained from subjects undergoing or considering IVF-related treatment.

A method of modifying a surface of a three-dimensional (3D) article includes immersing at least one part of the 3D article in a buffered solution of functionalized peptides, allowing reaction between the functionalized peptides and reactive groups on the surface of the 3D article; and washing the at least one part of the 3D article to remove unreacted functionalized peptides. The surface-modified 3D article includes a plurality of peptides covalently bonded to the surface of the 3D article via a cysteine bridge. The surface- modified 3D article can be used as a scaffold for the formation of biological tissue or bodily implants.

The present invention provides a production method for a three-dimensional muscle tissue including the steps of: preparing an approximately rectangular first cell module containing skeletal myoblasts in a hydrogel, and having a plurality of approximately rectangular holes parallel to each other, and an approximately rectangular second cell module containing skeletal myoblasts in a hydrogel, and having a plurality of approximately rectangular holes parallel to each other at positions different from those of the first cell module in a vertical direction; alternately stacking the prepared first cell module and the prepared second cell module to obtain a stack; subjecting the skeletal myoblasts contained in the obtained stack to proliferation culture; and inducing the proliferated skeletal myoblasts to differentiate into myotubes.

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 present disclosure relates to methods, products, and compositions useful for deriving bovine induced blastoids. Methods are described that allow for juvenile stem cell aggregation and induced blastocyst formation. The invention further provides an aggregation culture medium and an induced blastocyst-like culture medium which are suitable for promoting aggregation of the young stem cells and preparing the induced blastocyst-like. The bovine induced blastoids can be used in selection and breeding procedures, including the replication of pre-implant embryos with desired characteristics and in vitro breeding including a cross-generation breeding cycle from parent germ cells and / or embryos to progeny embryos.

The present disclosure relates to a transplantable cell therapy product composition for diabetes mellitus that contains an aggregate of insulin-secreting cells derived from stem cells. In the present disclosure, an NF microwell array membrane was fabricated by applying a molding process to an electrospun, permeable, biodegradable polycaprolactone (PCL) NF membrane and thus allows gases and soluble factors to permeate therethrough. The NF microwell of the present disclosure could provide more nutrients to the iPSC aggregates than conventional impermeable PDMS microwells, thus enhancing survival and differentiation capabilities of the cells. Additionally, the NF membrane was attached singly to the subcutaneous tissue and to the surface of organs such as liver and peritoneum without the need for a fixing material or separate sutures and was integrated with surrounding tissues, resulting in higher insulin secretion than PDMS microwells. Therefore, the present disclosure can be effectively utilized as a composition for the prevention or treatment of diabetes.

A cell culture microcarrier bead is proposed. The microcarrier bead comprises a bead body having its surface flecked with plasmonic nanoparticles. In a second aspect, the invention relates to a cell culture reactor, containing a cell culture medium and the proposed microcarrier beads. A third aspect of the invention concerns a method for observing living cells on such microcarrier beads. Yet a further aspect of the invention relates to a method for packing nanoparticles on a carrier body.

The invention provides a method for preparing an ordered cell-containing microarray, and a system for preparing an ordered cell-containing microarray.

The present invention provides a cardiac tissue model comprising cardiac tissue having at least one lumen or central cavity, wherein the cardiac tissue model comprises at least two different cardiac tissues selected from left ventricular tissue, right ventricular tissue, atrial tissue, outflow tract tissue, atrioventricular canal tissue, sinoatrial node tissue, and atrioventricular node tissue, the central cavity can be shared by at least two different cardiac tissues, and / or at least two different cardiac tissues comprise the ability to propagate calcium signaling connections and / or tissue contractions, a method of generating such a tissue model, and the use of the tissue model for screening purposes.

To provide a method for generating oligocortical spheroids (OCS) from pluripotent stem cells (PSC).SOLUTION: A method comprises the steps for: a) generating a neurocortical spheroid (NCS) through neurocortical patterning of pluripotent stem cells; b) subjecting the neurocortical spheroid to timed exposure to defined oligodendrocyte lineage growth factors and / or hormones, to promote proliferation, survival and / or expansion of native oligodendrocyte progenitor cell (OPC) populations within the neurocortical spheroid, thereby generating the oligocortical spheroid, wherein the oligocortical spheroid contains oligodendrocyte progenitor cells capable of differentiating into myelinating oligodendrocytes (ODCs) that are capable of myelinating axons.SELECTED DRAWING: Figure 1-1

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.

The present disclosure provides thymic cells, thymic organoids, and animal models comprising such thymic cells and thymic organoids. It also provides methods for inducing differentiation of pluripotent stem cells into definitive endoderm (DE), anterior foregut endoderm (AFE), ventral pharyngeal endoderm (VPE), thymic epithelial progenitor (TEP) cells, thymic epithelial cells (TECs) and thymic organoids in vitro and in vivo.

The present disclosure relates generally to methods and systems of producing Schwann cells from pluripotent stem cells under fully defined conditions. The Schwann cells produced by the disclosed methods find applications as models of the enteric nervous system, tools for high throughput screening of potential therapeutics for treatment of enteric neuropathies, and in regenerative medicine.

The invention relates to a polylactic acid microcarrier and a method for cell amplification by using the polylactic acid microcarrier. The preparation method disclosed by the invention is simple and convenient to operate, the prepared polylactic acid microcarrier is uniform in particle size and pore size distribution, the microcarrier is subjected to surface retreatment, more active sites are provided for cell adhesion, a three-dimensional matrix environment closer to a body is provided, cell amplification is facilitated, the dryness and differentiation potential of stem cells are maintained, and the preparation method is suitable for large-scale popularization and application. And the microcarrier can be repeatedly used, so that the use cost is reduced.

The present invention addresses the problem of providing a method for producing a suprachiasmatic nucleus cell organoid. The problem is solved by a method for producing a suprachiasmatic nucleus organoid comprising: (1) a step for culturing pluripotent stem cells in an intermediate medium between an undifferentiation maintenance medium and a nerve differentiation medium; (2) a step for culturing the cells obtained in step (1) in a nerve differentiation medium; and (3) a step for performing an operation for forming a cell mass prior to the start of step (2), wherein (A) the number of raw material cells seeded in order to form one cell mass is 6000 or more, and (B) when the start of step (2) is taken as day 0, the medium contains a sonic hedgehog signaling pathway agonist from day 3 onward.

The application provides an islet-vascular co-culture system based on a microfluidic chip, a building method and application, and the building method of the islet-vascular co-culture system comprises the following steps: S1, preparing microfibers loaded with islets; S2, preparing an assembled chip, the assembled chip comprises a containing cavity, first microchannel networks and second microchannel networks are symmetrically arranged on two sides of the containing cavity, and the first microchannel networks and the second microchannel networks are respectively communicated with a liquid inlet and a liquid outlet at an end away from the containing cavity; S3, controlling the flow of the liquid inlet to be 6-8 muL / min to carry out dynamic culture. The application can simulate the key structure and characteristics of islet tissue, can evaluate a diabetes treatment drug in vitro, can bridge the gap between in vitro and in vivo models, and can provide new tools and methods for the fields of drug development, disease model construction and personalized medical treatment.

The invention belongs to the cross technical field of veterinary medicine and biomedicine, and relates to a construction and evaluation method of a dog spontaneous breast cancer organoid model, which specifically comprises the following steps: (1) obtaining dog spontaneous breast cancer tissues through operation or biopsy, separating tumor tissues under sterile conditions, and carrying out fine enzymolysis digestion treatment to obtain dog spontaneous breast cancer tissues; obtaining a single-cell suspension; and (2) inoculating the single-cell suspension into a three-dimensional culture system, carrying out three-dimensional culture by adopting a culture medium containing EGF, FGF and B27 in combination with a growth factor, promoting the growth and amplification of the breast cancer organs, and carrying out timely passage according to the growth condition so as to maintain the activity of the model. The model constructed by the invention completely retains genetic heterogeneity and pathological microenvironment of primary tumors, does not need gene editing, is simple and convenient to operate and low in cost, is particularly suitable for individual drug sensitivity detection of pet dogs, and provides an efficient and high-simulation experimental platform for dog breast cancer mechanism research and clinical precise medication.