296results about "Respiratory/lung cells" patented technology

The present invention uses ex vivo human airway cultures to assess the human transmissibility and replication competence of influenza and coronavirus strains. By comparing pandemic influenza A subtype H1N1 and highly pathogenic avian influenza H5N1 as reference strains, the transmissibility risk of various viruses was evaluated and categorized. Additionally, an in vitro model evaluated virus-induced impairment of alveolar fluid clearance (AFC) as an indicator of disease severity. The study revealed correlations between bronchus viral replication, human transmission, AFC impairment, and clinical disease severity across different influenza and coronavirus strains.

The invention belongs to the technical field of biology, and particularly relates to a human metapneumovirus mouse adapted strain and application thereof. Compared with a wild strain, the strain has higher susceptibility to animals; the strain can be used for treating BALB / c mice and can cause typical clinical symptoms and even death, for example, after the strain infects the BALB / c mice, the disease symptoms are obvious, the weight of 83% infected mice is severely reduced to 75% or below and reaches the humanity killing standard, and the death rate reaches 50% or above 4 days after the strain infects the BALB / c mice; the strain can be used for constructing a cell model or an animal model of human metapneumovirus infection or diseases caused by the human metapneumovirus infection, and is further used for screening drugs for preventing and / or treating the human metapneumovirus infection or the diseases caused by the human metapneumovirus infection. In addition, the strain can also be used for preparing medicines for preventing and / or treating human metapneumovirus infection or diseases caused by the human metapneumovirus infection, preparing antibodies or antiserum for resisting the human metapneumovirus and the like.

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 present disclosure relates to a method for preparing a respiratory organoid by proliferating and differentiating respiratory epithelial cells. Unlike the conventional respiratory organoids in which epithelial cells existing in the apical region of the respiratory organ are generated on the interior of the organoid, the respiratory organoid of the present disclosure is shown in a form in which ciliated structures, which are epithelial cells, are located in the outer layer of the organoid, secretory cells are present on the interior, and basal stem cells existing at the base of respiratory tissue are also present on the interior of the organoid. It has a form similar to an actual respiratory tissue, and it was observed that the epithelial cells exhibited normal function, such as motility of ciliated cells. Therefore, when an organoid is prepared using the preparation method of the present disclosure, a respiratory organoid that can well mimic actual tissue can be prepared.

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 invention belongs to the technical field of biological medicine manufacturing, and particularly provides a preparation method of an engineered exosome capable of loading mitochondria for treating pulmonary fibrosis and the engineered exosome. The method comprises the following steps: constructing fusion plasmids for expressing mitochondrial targeting peptide fragment-therapeutic protein genes, dissolving the fusion plasmids in non-enzyme ultrapure water, adding the fusion plasmids into a lipid nano-preparation, and fully mixing to obtain a nano-preparation containing the therapeutic protein genes; the nano preparation is delivered into alveolar epithelial cells and is efficiently transfected, so that the homeostatic balance of the injured alveolar epithelial cells in the development of pulmonary fibrosis is effectively regulated and controlled, the secretion of an exosome with a therapeutic effect is promoted, and the engineered exosome is further obtained through mannose modification. The exosome can effectively load mitochondria and sequentially and accurately target macrophage mitochondria on the basis of a mannose target head and a mitochondrial targeting peptide fragment, so that macrophage mitochondrial homeostatic balance is effectively regulated and controlled, the purpose of reverse treatment of pulmonary fibrosis is achieved, and the exosome has high clinical application value.

The invention provides a lung stem cell peptide for treating pulmonary nodule as well as a preparation method and application thereof, and belongs to the technical field of biological pharmacy. The method comprises the following steps: performing induced differentiation on iPSC to obtain lung stem cells; lung stem cells are cultured and centrifuged, supernatant is collected, and a lung stem cell crude solution is obtained; mixing the lung stem cell crude liquid with protease, performing enzymolysis, centrifuging, concentrating, and performing spray drying to obtain the lung stem cell peptide for treating pulmonary nodule. By optimizing the composition of each culture medium, the iPSC is induced and differentiated into the lung stem cells, then the lung stem cell peptide is extracted and separated from the lung stem cells, and the extracted lung stem cell peptide migrates to the lung in a vein reinfusion mode, so that the generation of lung tumor nodules can be effectively inhibited, the number of the lung tumor nodules is greatly reduced, and the lung tumor nodules can be effectively inhibited. Toxic and side effects and adverse reactions are avoided.

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 application discloses a combined drug for treating lung cancer. It is found for the first time that lomethixol can be used alone or in combination with other drugs to have a significant anticancer effect on NSCLC patients and NSCLC patients resistant to osimertinib, and in addition, it is found that lomethixol is a potential autophagy inducer and can activate cell autophagy. The application provides a new clinical treatment method for NSCLC patients and NSCLC patients resistant to osimertinib, and has a wide application prospect.

The invention discloses an inflammatory pulmonary epithelium injury treatment evaluation method based on alveolar organs, and relates to the technical field of medical biology. The invention mainly aims at alveolar epithelium injury and repair in inflammatory lung injury, and establishes an organ-like model for simulating alveolar epithelium injury. By using the model, the efficacy of related drugs on alveolar epithelium injury repair can be detected in a targeted manner, and an efficient drug screening and evaluation platform and method are provided for pulmonary epithelium repair in inflammatory lung injury. According to the technical scheme, the method comprises the steps that mouse alveolar epithelial stem cells are separated through magnetic bead sorting, an alveolar organoid inflammatory lung injury model is established through LPS exposure, alveolar organoid is treated through drugs, the organoid generation number, the organoid formation rate, the epithelial cell proliferation rate and the AT2-AT1 differentiation efficiency are detected, and the drug treatment effect is evaluated.

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.

Provided herein are artificial lung organoids. The artificial lung organoids may include an epithelial cell layer comprising mammalian lung epithelial cells, a stromal cell layer comprising mammalian lung fibroblast cells and an endothelial cell layer comprising mammalian endothelial cells. The artificial lung organoids may optionally include a porous membrane between said epithelial cell layer and said stromal cell layer and / or between said stromal cell layer and said endothelial lung cell layer.

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 discloses application of NCCRP1 as a lung adenocarcinoma immunotherapy biomarker, and belongs to the field of biological medicine. Bioinformatics analysis finds that the NCCRP1 is remarkably and highly expressed in lung adenocarcinoma tissues, and the expression level of the NCCRP1 is in negative correlation with prognosis of a patient. Secondly, the expression level of lung adenocarcinoma cells NCCRP1 is in negative correlation with infiltration of anti-tumor immune cells such as CD8 + T cells. Experiments show that the knockout of the NCCRP1 can significantly inhibit the growth of mouse lung tumors by increasing the infiltration quantity of CD8 + T cells and NK cells in a lung adenocarcinoma microenvironment. The invention provides a new target spot for immunotherapy of lung adenocarcinoma, including NCCRP1 gene editing (knockout / knockdown) and application of an inhibitor in preparation of antitumor drugs.

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.

The invention uses ex vivo human airway cultures to assess the transmissibility and replication ability of influenza and coronavirus strains. By comparing the pandemic influenza A H1N1 and the highly pathogenic influenza A H5N1 as reference virus strains, the spreading risks of various viruses are evaluated and classified. In addition, an in vitro model is used to assess the degree of alveolar fluid clearance (AFC) impairment caused by the virus as an indicator of the severity of the disease. The research reveals the relevance among the virus replication ability, human transmission, AFC damage degree and clinical disease severity of different influenza and coronavirus strains in bronchus.

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 a primary cell culture medium for culturing primary lung cancer epithelial cells, which contains at least one of an MST1 / 2 kinase inhibitor, a ROCK kinase inhibitor, a fibroblast growth factor 7, a B27 supplement, and a N2 supplement, a hepatocyte growth factor, an insulin-like growth factor 1, an interleukin 6, and a TGFβ type I receptor inhibitor. The present invention also relates to a culturing method using the primary cell culture medium and its application in drug efficacy evaluation and screening. The culturing method uses the above-mentioned primary cell culture medium to culture primary cells on a culture vessel coated with extracellular matrix glue, so that the primary cells proliferate rapidly. The cell model obtained by the primary cell culture medium and the primary cell culturing method of the present invention can be used for drug efficacy evaluation and screening.