170results about "Respiratory/lung cells" patented technology

The invention discloses a mouse lung organ and fibroblast co-culture method, which comprises the following steps of: performing in-vitro mouse lung organ culture on mouse lung tissue primary epithelial stem cell spheres (pneumospheroids) in a Transwell upper chamber, and after the lung organs in the Transwell upper chamber are differentiated, replacing with a lower chamber paved with mouse lung fibroblasts, therefore, a gas-liquid interface co-culture system of the mouse lung organ (upper chamber) and the fibroblast (lower chamber) is formed, and the hierarchical structure, morphological characteristics and regeneration function of the upper chamber organ and the phenotype, function and related signal path activation level of the lower chamber fibroblast can be dynamically researched.

The invention relates to the technical field of biological medicine, in particular to a drug screening model construction method based on airway organs, a model and application. The construction method of the drug screening model of the airway organoid comprises the steps of obtaining 2D airway epithelial cells, culturing the airway organoid, establishing an airway organoid inflammation model and / or establishing an airway organoid mucus secretion / goblet cell metaplasia model. Wherein the model is established according to the culture mode of the airway organ in cooperation with a specific stimulant for proliferation and differentiation. Compared with a traditional animal model, the organ-like model is low in cost and short in construction period, can better simulate the pathological process, can realize airway organ-like culture and establishment of an inflammation model and a mucus secretion / goblet cell metaplasia model at the same time, and has a good application prospect in drug detection of asthma and COPD diseases.

The invention relates to a lung cancer organoid and peripheral blood source immune cell co-culture model and a construction method thereof. Specifically, the invention provides a construction method of a macrophage and tumor organoid co-culture model for evaluating anti-tumor activity, monocytes of autologous or allogeneic peripheral blood of a patient are induced and differentiated into high-purity macrophages in vitro, and the high-purity macrophages and lung cancer tumor organoid are subjected to three-dimensional co-culture in matrigel; the model aims at highly reducing a core interaction network of tumor cells and macrophages in TME; the dynamic change of the polarization state of the macrophage is simulated and observed; the problem that an organ-like model is incomplete due to immune component deficiency or spatial positioning distortion of an existing model is effectively solved, an experimental platform closer to the physiological state is provided for tumor immune microenvironment research, and therefore development of the tumor immune treatment field is promoted.

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.

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.

This invention provides a lung stem cell peptide for treating pulmonary nodules, its preparation method, and its application, belonging to the field of biopharmaceutical technology. The invention involves inducing iPSCs to differentiate into lung stem cells; culturing the lung stem cells, centrifuging, collecting the supernatant to obtain crude lung stem cell solution; mixing the crude lung stem cell solution with a protease for enzymatic hydrolysis, centrifuging, concentrating, and spray-drying to obtain the lung stem cell peptide for treating pulmonary nodules. This invention optimizes the composition of each culture medium to induce iPSCs to differentiate into lung stem cells, then extracts and separates lung stem cell peptides from these cells. The extracted lung stem cell peptides are intravenously reinfused into the lungs, effectively inhibiting the formation of pulmonary tumor nodules, significantly reducing the number of pulmonary tumor nodules, and without toxic side effects or adverse reactions.

A method of testing a test substance in an in vitro model of a human tracheobronchial respiratory tract, includes: providing the in vitro model including a cell culture including airway epithelial cells (AECs), a basolateral compartment below the AECs and an apical compartment above the AECs, wherein the AECs form a barrier between the basolateral AND apical compartments; adding the test substance to the apical and / or basolateral compartment; adding a tracer to the basolateral compartment, which is fluorescent, has a molecular weight within 5 kDa of human albumin and is added before, during or after adding the test substance; incubating the cell culture system in a presence of the tracer; collecting at least one sample from the apical compartment; and measuring a fluorescence thereof to determine an effect of the test substance on the AECs. A kit is also disclosed.

Methods and systems are described for a high-level approach to organ generation by self-assembly using bioprinting, overcoming longstanding challenges in replicating the intricate architecture and function of complex organs. In some embodiments, these methods are used to produce lung tissues and microtissues, demonstrating how mini-lung constructs can be produced on a small scale through specialized processes and media. In such embodiments, methods and systems for generating bioprinted lung tissues and microtissues are provided, as are tissues made by such methods. The tissues find use in implantation, drug discovery, personalized medicine, and other applications. Methods for the large-scale manufacturing of lung epithelial stem cells are also provided.

The purpose of the present invention is to provide a novel method for culturing tissue-derived cells. The method uses a cell culture method including a step for co-culturing cells derived from the skin and at least one type of cell selected from among cells derived from the intestine, cells derived from the heart, cells derived from the brain, cells derived from the stomach, cells derived from the embryonic membrane, cells derived from the kidney, and cells derived from the lung. And culturing the skin-derived cells.

The application belongs to the technical field of biotechnology, and particularly relates to a human metapneumovirus mouse-adapted strain and application thereof. Compared with wild strains, the strain can exhibit stronger susceptibility to animals, and can cause typical clinical symptoms and even death. For example, after the strain infects BALB / c mice, the disease symptoms are obvious, 83% of the mice have a severe decrease in body weight to below 75% after infection, and have reached the humane killing standard. Four days after the strain infects BALB / c mice, the mortality rate is above 50%. The strain can be used to construct a cell model or an animal model of human metapneumovirus infection or a disease caused by the human metapneumovirus, and is further used to screen drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus. In addition, the strain can also be used to prepare drugs for preventing and / or treating human metapneumovirus infection or a disease caused by the human metapneumovirus, and to prepare antibodies or antisera against the human metapneumovirus.

The present disclosure provides a method for producing lung mesenchymal cells that can be used as feeder cells in induction of alveolar epithelial cells. A method for producing lung mesenchymal cells of the present disclosure includes culturing mesodermal cells in a presence of a mesenchymal cell-inducing factor, KGF, and FGF 10 so as to induce differentiation of the mesodermal cells into lung mesenchymal cells.

The invention belongs to the technical field of bioengineering, and relates to heparan sulfate defect type matrigel as well as a preparation method and application thereof. The preparation method comprises the following steps: providing natural matrigel; providing immobilized heparinase, wherein the immobilized heparinase can specifically catalyze and degrade heparan sulfate in the natural matrigel; carrying out contact reaction on the natural matrigel and the immobilized heparinase; and after the reaction is finished, separating the immobilized heparinase. According to the preparation method, the characteristics that the immobilized enzyme is convenient to separate and recycle are utilized, the problems that in a traditional solution enzyme method, enzyme residues exist, the reaction is difficult to accurately control, and subsequent purification steps are complex are solved, and a standardized heparan sulfate defective matrigel product which is controllable in component, free of residual enzyme and high in stability is provided. The method has wide application value in the fields of tumor mechanism research, anti-tumor drug screening, tissue engineering and the like.

The invention relates to a thermostable FGF10 polypeptide possessing FGF10 activity and having or comprising at least 85% sequence identity to SEQ ID NO:3 that has or comprises an amino acid sequence from Ser69 to Ser208 of SEQ ID NO:1, or to SEQ ID NO:5 that has or comprises an amino acid sequence from Leu40 to Ser208 of SEQ ID NO:1, or the fragments thereof, comprising at least an amino acid substitution L152F. Preferably, any of amino acid substitutions V123I, Q175E, and N181D can be further included. The invention further discloses the use of subjected thermostable FGF10 polypeptides in regenerative medicine or other related medical applications or cosmetics. Further, it discloses a culture medium comprising subjected thermostable FGF10 polypeptides suitable for proliferation and differentiation of the human embryonic stem cells or formation and differentiation of spheroids and organoids.

The invention provides a construction method of a non-small cell lung cancer histone H1.3 arginine methylation point mutation cell model. The construction method comprises the following steps: designing mutation primers H1.3 R80A-F and H1.3 R80A-R; the method comprises the following steps: by taking a pCDH-HA-H1.3-Flag plasmid as a template, carrying out PCR (Polymerase Chain Reaction) amplification by adopting a mutation primer, digesting a product by DMT enzyme, and converting a competent cell to obtain a mutant plasmid; and co-transfecting the mutant plasmid and a helper plasmid to a packaging cell, collecting a virus solution, filtering, infecting an A549 cell, adding Polybrene to assist infection, culturing, and screening a stably transfected cell strain by using puromycin to obtain the recombinant plasmid. The invention also provides application of the mutant cell model obtained by the construction method. According to the invention, the blank of histone H1.3 methylation research is filled, the 80th arginine is clear as a core modification site, and the established model provides a new tool for lung cancer mechanism research and drug research and development.

The invention relates to the field of biological medicines, and discloses application of a GOSR1 gene in prevention and / or treatment of respiratory pathogens. The GOSR1 gene is specifically knocked down, so that the gene and protein expression level of the pathogen can be reduced, and the proliferation of the respiratory pathogen can be remarkably inhibited. In addition, the invention also discloses that overexpression of the GOSR1 gene can activate proliferation of pathogens in cells infected by respiratory pathogens. Based on the regulation effect of the GOSR1 gene on respiratory tract pathogen proliferation, a brand new strategy is provided for treatment of respiratory tract pathogen infection, and the application is particularly suitable for intervention of influenza A virus infection.

The present invention relates to a preparation method of a composition for culturing lung organoids, a composition thereby, and an organoid culture method using the same. The present invention is capable of creating an environment that is more similar to an actual tissue than a conventional MATRIGEL®-based culture system, and in particular, exhibits an effect of facilitating tissue differentiation in lung organoid culture and effectively developing into a form that is similar to an actual tissue.

Provided herein are lung forming progenitor cell therapies which do not require pre-conditioning treatments. Administration of the progenitor cells can replace or replenish populations of host-derived patch forming cells to reverse or inhibit fibrosis, promote healing, and improve lung function. Further disclosed herein are methods for monitoring fibrosis, and for determining whether a subject will be receptive to progenitor cell treatments.

The invention provides a lung-kidney interaction simulation system based on an organ chip, and the system comprises a lung chip module which is used for simulating a qi-blood barrier structure of a lung, and the lung chip module is inoculated with human alveolar epithelial cells and human lung microvascular endothelial cells; the kidney chip module is used for simulating a capillary lumen of a kidney tubule and a kidney tubule lumen structure, and human kidney tubule epithelial cells and human umbilical vein endothelial cells are inoculated on the kidney chip module; the connecting channel is used for communicating the lung chip module with the kidney chip module and allowing the culture medium to flow between the lung chip module and the kidney chip module; and wherein a klebsiella pneumoniae liquid for causing lung injury is applied to the upper layer of the lung chip module. The invention provides a novel simulation system for the research of lung-kidney interaction, and solves the limitation of animal experiments.

The invention belongs to the technical field of cell infection, and particularly relates to an organoid infection method based on microinjection. The objective of the invention is to accurately inject fluorescently-labeled pathogens into the interior of the organoid, so that the physical barrier of matrigel is overcome, and meanwhile, the physiological structure and function of the organoid are reserved to the greatest extent. Specifically, fixed-point infection of pathogens in the organoid is realized by utilizing a microinjection method, and the pathogens can effectively bypass a matrigel barrier, so that targeted infection of a specific region in the organoid is realized, and the infection efficiency and specificity are remarkably improved; meanwhile, matrigel does not need to be digested, the complete three-dimensional structure, cell polarity and intercellular connection of the organoid can be reserved to the maximum extent, the infection model is closer to the real physiological state in the body, and therefore the reliability of the result is improved.

The present invention provides processes for producing a bioengineered lung (BEL) from an acellular lung matrix that has been treated with growth hormones, seeded with primary lung cells, and cultured in a bioreactor. Also provided are BELs and methods of transplanting the BEL into a subject in need of a lung transplant, and methods for using BELs for the study of the lung microbiome and its role in lung development and remodeling.

The field of the invention is in vitro assays for the production of in vitro organ culture systems and the use of such in vitro organ culture systems for screening assays in drug screening, patient selection and personalized medical context. More specifically, the present invention relates to an in vitro method of establishing a tumor organoid from a cancer tissue sample obtained from a subject, an in vitro method of providing immune cells activated by an autologous tumor organoid, a method of preparing an in vitro organ culture system for mimicking interaction of a tumor with an immune system of a subject, and a method of preparing an in vitro organ culture system for mimicking interaction of a tumor with an immune system of a subject. The invention relates to an in vitro organ culture system, and to an in vitro organ culture system produced thereby, to a method for determining the reactivity of a tumor to at least one immunooncology drug therapy using said in vitro organ culture system, and to a kit for preparing an in vitro organ culture system that mimics the interaction of a tumor with the immune system of a subject, as disclosed herein.

The invention belongs to the technical field of gene engineering, and particularly relates to a construction method and application of a recombinant vector based on bCoro1a gene 3 'UTR editing. The recombinant vector consists of an upstream homologous arm of a targeting site, 3 'UTR inserted into a target sequence, 200bp of a nucleotide tailing signal of bCoro1a, a selection marker and a downstream homologous arm of the targeting site; the targeting site is the 29 or 70 site of the 3 'UTR of the bCoro1a gene; the sequence of the 3 'UTR of the bCoro1a gene is as shown in SEQ ID NO. 48; the selection marker comprises enhanced green fluorescent protein and puromycin, the bCoro1a 3 'UTR edited gene expression regulation and control system is based on bCoro1a 3' UTR edited gene expression regulation and control system, and the expression level of bCoronin-1A is reduced in a targeted manner by inserting a specific bta-miR-27b target sequence into 3 'UTR, so that M.tb immune escape is inhibited, and the M.tb infection resistance of a host is improved.

We provide cultured cell sheets. [Solution] A cultured cell sheet is provided, which is made from cells collected from nasal mucosal tissue, and contains 50-90% undifferentiated cells relative to the total number of cells, wherein, in one embodiment, the undifferentiated cells are positive for p63, and the cultured cell sheet is a mixture of nasal mucosal epithelial cells and nasal mucosal epithelial stem cells or nasal mucosal epithelial progenitor cells, as well as one or more of the following cells: other epithelial cells or other epithelial stem cells, mesenchymal stem cells, fibroblasts, vascular endothelial cells, vascular endothelial progenitor cells, and adipocytes. [Effects] The cultured cell sheet of the present invention engrafts on areas of mucosal tissue defects such as inflammation and granulation tissue formation on the surface of bone tissue in the middle ear, promoting the regeneration of mucosal tissue, including mucosal tissue covering bone tissue, efficiently suppressing inflammation occurring in the bone tissue of the middle ear, and inhibiting characteristic fibrosis, granulation tissue formation, and poor epithelial formation that occur during bone tissue inflammation.

The present disclosure aims to provide a structure that allows access to the lumen of a cellular structure. The present disclosure provides a structure including: a sac-like cellular structure; and a tubular body, wherein the cellular structure includes a lumen and a cell layer at least partially covering the lumen, the cell layer is composed of cells, and the tubular body is configured to allow the lumen of the cellular structure to be in communication with an outside of the cellular structure.

An object of the present invention is to provide a cultured cell sheet that can solve problems accompanied by defects of mucosal tissue as described above, such as inflammation in mucosal tissues covering bone tissues and granulation tissue formation. In order to solve the problems described above, the present inventors have conducted research and development with examinations from various angles. As a result, it has been found that a cultured cell sheet obtained by culturing cells collected from a nasal mucosal tissue as a source under specific conditions, and reconstructing the cells into a sheet are engrafted onto mucosal tissue defective sites, such as due to inflammation on the surface of middle ear bone tissues and granulation tissue formation; promote the regeneration of mucosal tissues, such as those covering bone tissues; efficiently reduce inflammation that occurs in a middle ear bone tissue; and prevent fibrosis, granulation tissue formation, defective epithelial formation, and the like that are characteristic of bone tissue inflammation, thereby demonstrating that the cultured cell sheet can solve the above-described problems. It is also found that a cultured cell sheet made from cells collected from a nasal mucosal tissue of the present invention is also useful for preventing inflammation of bone tissues in other areas of the body other than the middle ear.

The invention provides a lung squamous cell carcinoma organoid culture medium and a culture method. The lung squamous cell carcinoma organoid culture medium is prepared from the following components: R-spondin-1, Noggin, EGF (Epidermal Growth Factor), FGF10 (Fibroblast Growth Factor), FGF7 (Fibroblast Growth Factor), A83-01, Y-27632, Nicotinamide, Insulin, B27 Supplement, N-Acetylcysteine, Alpisib, GlutaMAX, primary cell antibiotics and the like. The lung squamous cell carcinoma organoid culture medium can be used for efficiently culturing lung squamous cell carcinoma tissues and successfully obtaining organoid, the formed organoid is highly similar to primary lung squamous cell carcinoma tissues in cell type, polarity and spatial hierarchical structure, and the micropathologic characteristics of tumors are truly reproduced. And the organoid can be stably passaged for a long time, so that an ideal and reliable preclinical model is provided for pathogenesis research, personalized drug screening and novel therapy development of lung squamous cell carcinoma.

The invention provides a construction method and application of a sepsis model based on a lung and kidney organ chip. The method comprises the following steps: S1, carrying out ultraviolet sterilization on an organ chip device; s2, modifying a transwell plug-in unit; s3, inoculating human lung microvascular endothelial cells and human umbilical vein endothelial cells; S4, inoculating human alveolar epithelial cells and human renal tubular epithelial cells; s5, transwell inserts are arranged in two holes in the middle of the organ chip device respectively, a mixed culture medium is added into liquid adding holes in the two sides of the organ chip device, and the lung chip and the kidney chip are communicated through a lower channel of the organ chip device; s6, dynamically culturing the lung and kidney organ chip; and S7, sucking up the culture medium in the constructed lung and kidney organ chip, adding the LPS-containing mixed culture medium into a lower channel, and respectively adding the LPS-containing mixed culture medium into the human alveolar epithelial cell side and the human renal tubular epithelial cell side of the lung and kidney organ chip and the transwell plug-in hole for culturing for 2 days. According to the invention, the lung and kidney organ chip is applied to the sepsis model for the first time, and the constructed sepsis model is more suitable for basic research and drug development of sepsis.

The present invention provides compositions and methods of preparing airway cells. In one aspect, an epithelial airway cell derived from an induced pluripotent stem (iPS) cell characterized by expression of airway cell surface markers and an ability to proliferate is described. In another aspect, methods of differentiating an iPS into an epithelial airway cell is provided. Engineered lungs, methods of making such engineered lungs comprising the epithelial airway cells and treating respiratory disorders are also disclosed.

Disclosed are methods, media, supplements and kits for expanding, maintaining, or differentiating mammalian epithelial cells, such as alveolar epithelial cells - AT1 and AT2 cells, in monolayer. The methods of this disclosure can be used to expand, maintain or differentiate mammalian alveolar epithelial cells in a 2D environment when such cells are seeded in the presence of one or more extracellular matrix proteins.

It is provided a protocol to induce the formation of lower respiratory tract organoids in only 21 days, the organoids containing markers of both proximal and distal regions. The organoids is obtained by leveraging cellular interactions between airway bronchial epithelial cells and fibroblasts. It is also provided the use of the organoids to study respiratory viral infections with strong similarity to adult lung tissue.