209 results about "Disease modelling" patented technology

The invention provides a method for drug screening and disease model construction of liver organoid, relates to the technical field of medical research, and adopts multi-modal data acquisition, fusion and intelligent analysis and quantum computing processing to optimize comprehensive application of data, so that the richness, accuracy and analysis efficiency of the data are remarkably improved, and the risk of drug screening and disease model construction of liver organoid is reduced. Multi-source data of genomics, transcriptomics and the like are collected comprehensively, an accurate research system of immune-hepatic disease interaction based on organoid is constructed, an in-vivo complex immune-hepatic disease microenvironment can be highly simulated, the proportion and activation state of immune cells are accurately regulated and controlled in an in-vitro model, functional and phenotypic changes of the immune cells are monitored in real time, and the immune-hepatic disease interaction accuracy is improved. According to the present invention, the interaction mechanism between the immune cells and the liver organ cells is deeply revealed, such that the accurate information basis is provided for the accurate determination of the drug action target, the development of the innovative drug for the immune check point or tumor microenvironment regulation is easily achieved, and the pertinence and the success rate of the drug research and development are substantially improved.

The invention provides a neuromuscular multi-element coupled organoid micro-fluidic chip, a device and a method. Relates to the technical field of organoid micro-fluidic chips. A first culture chamber, a second culture chamber and a third culture chamber are arranged in a culture layer of the chip; the first culture room is used for culturing nerve cells; the second culture room is used for culturing muscle cells; a microfluidic channel for communicating the first culture chamber with the second culture chamber is arranged between the first culture chamber and the second culture chamber; the third culture room is arranged on the micro-fluidic channel and is used for culturing a neuromuscular joint formed by convergence of axons of nerve cells and muscle cells; a microelectrode array plate is arranged in the circuit layer and is used for performing electrical stimulation and electrophysiological function detection on neuromuscular cells. Through the chip, co-culture of in-vitro nerves and muscle organs can be realized, a complex interaction relationship in a neuromuscular system can be accurately simulated, and the chip can also be used for constructing various neuromuscular related disease models.

The invention belongs to the technical field of organoid chips of biological medicine, and particularly relates to a multi-organoid chip and a preparation method thereof. The multi-class organ chip comprises an upper-layer chip and a lower-layer chip which are detachably arranged; 2-10 organ-like chambers are embedded in the upper-layer chip, and 2-5 porous membranes are arranged at the bottoms of the organ-like chambers; the lower-layer chip is provided with a flow channel, and the flow channel is communicated with the organ-like chamber. Compared with the existing multi-class organ chip technology, the multi-barrier construction in the independent cavity is realized, and meanwhile, the multi-class organ chip is combined with a transwell component for use, so that the personalized combination of the multi-class organ chips is realized. The method is economical and fast, can play an important role in promoting the fields of precise medicine, medicine evaluation, disease models, especially in-vitro evaluation and simulation of traditional Chinese medicine and the like, and is suitable for popularization and application.

The invention discloses a Parkinson's disease animal model, a construction method and application, and relates to the field of animal genetic engineering and disease model construction. According to the model, seven mutation sites related to the human familial Parkinson's disease are introduced behind an initiation codon of a fourth exon of an endogenous SNCA gene, and the mutation sites are A18T, A29S, A30P, E46K, H50Q, G51D and A53T and have genotypes related to the human familial Parkinson's disease. The pig model shows nigra dopaminergic neuron reduction and cortical neuron reduction in the newborn period, and the core pathological process of the human Parkinson's disease can be systematically reproduced. The model can be widely applied to Parkinson's disease pathogenesis research, drug screening and cell therapy effect evaluation, and has important scientific research and preclinical application values.

The construction of a non-human primate Usher syndrome disease model having USH2A gene mutation, wherein a target sequence against the mRNA of the non-human primate USH2A gene is established and a corresponding sgRNA is synthesized, then a knockout vector is constructed, the sgRNA and a Cas9 mRNA are mixed and injected into a non-human primate embryo to obtain a non-human primate embryo having USH2A gene knockout, and then by means of embryo transfer, a non-human primate having USH2A gene knockout is obtained, which exhibits symptoms of Usher syndrome. Upon detection and identification of the knockout effect, the knockout of the USH2A gene can be effectively realized in a non-human primate cynomolgus monkey embryo and cynomolgus monkeys, which provides a new and effective disease model for the diagnosis and treatment of Usher syndrome in non-human primates.

The invention provides preparation, sorting and application of LGE / CGE organoid and GABAergic neurons, and belongs to the field of brain organoid differentiation and GABAergic neurons. The preparation method of the LGE / CGE type organ comprises the following steps: S1, performing embryoid formation and initial culture on the 0th-4th day; s2, performing nerve induction and matrigel embedding on the 5th-11th day; s3, culturing the brain organoid on the 12th day to the 19th day; s4, dynamically culturing and screening the brain organoid on the 20th-35th day; and S5, carrying out differentiation and collection on LCOs of the LGE / CGE type organs after the 35th day. According to the method, the LGE / CGE organs containing the high-purity GABA energy neurons are obtained through differentiation from the brain organs for the first time, the high-purity LGE type GABA energy neurons and the high-purity CGE type GABA energy neurons are obtained through a specific sorting method, and the method has the deep application potential in the fields of disease model construction, medicine research and development and the like.

The invention provides a human-derived recessive retinal degeneration disease organ-like model based on Prom1 gene knockout. Specifically, the invention provides a specific gRNA targeting a Prom1 gene exon 9, and the specific gRNA can be used for efficiently knocking out the Prom1 gene of the human embryonic stem cell. The invention also provides a Prom1 gene knockout stem cell line derived from the human embryo pluripotent stem cell H9 and a retinal organ disease model. The retina-like organ can provide a research model for clinical treatment drugs.

The application discloses a method for constructing a mouse model of atherosclerosis induced without high-fat diet, and relates to the technical field of disease model construction.The mouse model construction method provided by the application obtains a mouse model that spontaneously exhibits atherosclerosis symptoms without high-fat diet induction.The mouse model of atherosclerosis induced without high-fat diet can be used for researching the pathogenesis of atherosclerosis and screening and evaluating drugs for atherosclerosis.

The invention belongs to the technical field of organoid chips of biological medicine, and particularly relates to an organoid chip for time sequence analysis and a preparation method of the organoid chip. The organ-like chip comprises an upper-layer chip and a lower-layer chip which are detachably arranged; the upper-layer chip is provided with a flow channel inlet, a flow channel and a flow channel outlet; the lower-layer chip is provided with organoid culture holes; and the flow channel inlet, the organoid culture hole, the flow channel and the flow channel outlet are communicated with one another. Compared with the existing organoid chip technology, the method has the advantages that the construction of the multi-channel independent cavities is realized, the culture of the organoid in each cavity in the same environment can be realized, the other cavities are not interfered when the organoid in a certain cavity is independently extracted, the organoid treated at different time is allowed to be evaluated, and the method is economical, rapid and high in efficiency. The method can play an important role in promoting the fields of precise medicine, medicine evaluation, disease models, especially traditional Chinese medicine in-vitro evaluation simulation and the like, and is suitable for popularization and application.

Described herein is a microphysiological system for models of disease. Specifically, induced pluripotent stem cells (iPSCs) and iPSC-derived cells, including those obtained from disease patients, are seeded onto microfluidic “chip” devices to study cellular development and disease pathogenesis. Herein, neurodegenerative disease modeling, including Parkinson's Disease (PD) is shown to reproduce key PD pathology in a vascularized human model that contains neurons relating to PD pathology. Such compositions and methods are used for research for PD biomarkers, patient screening for PD risk assessment, and therapeutic discovery and testing. A panel of biomarkers are generated through analysis of living PD-chips by neural activity, whole transcriptomic, proteomic, and metabolomic analysis, and functional enzyme tests of media and tissue. Introducing therapeutics through a vasculature channel, coupled with blood brain barrier penetration studies can be assessed for efficacy in the human neural cells present in the PD-Chip.

The invention relates to the field of medical biology, and discloses a construction method and device of a bipolar disorder experimental animal model and application, the construction method comprises the following steps: selecting a healthy experimental animal to carry out environmental adaptation feeding before modeling, then carrying out sleep deprivation stimulation for 3-5 days continuously, and carrying out sleep deprivation stimulation for 15-18 hours continuously and 6-9 hours for rest every day so as to freely eat and feed water; and after the sleep deprivation stimulation is finished, evaluating the surface validity of the disease model for the experimental animal by adopting behavioral test. The construction method provided by the invention is simple to operate and easy to repeat, can better simulate the disease of a clinical bipolar disorder patient, and provides a new model normal form for pathogenesis research and clinical drug screening of the bipolar disorder, and the disease model can be maintained for a long time.

The invention belongs to the technical field of animal model construction, and particularly relates to a non-alcoholic fatty liver disease mouse model based on humanized PNPLA3 I148M transgene as well as a construction method and application thereof, the construction method comprises the following steps: step i: designing CDS of a human gene and a key 3 'UTR element for patent medicine of a small nucleic acid drug in gene plasmid DNA of the constructed model; step ii, using Luc2 or iRFP to report gene expression in the gene plasmid DNA for constructing the model; and (iii) a disease model of the fatty liver disease induced by combining plasmid DNA transient transfer with high-fat diet. The mouse model constructed by the invention has the advantages that a specific exogenous gene expression signal of the liver is detected in real time through small animal living imaging, a human PNPLA3 protein is detected and overexpressed through serum ELISA, PNPLA3 I148M mRNA is detected and overexpressed through liver RT-qPCR, the phenotype of the non-alcoholic fatty liver disease of the liver is determined through 15-week-old pathology, and the whole process of modeling for 8 weeks does not need to be operated in a biosafety secondary laboratory.

The invention relates to the field of biology, in particular to a fluorescence and bar code pedigree tracking alveolar organ and a construction method and application thereof. The invention particularly aims at the problems of specific sorting of AT2 cells, culture of organoids and pedigree retracking in the disease process, and is used for simulating and researching the in-vitro pathological process of chronic obstructive pulmonary disease (COPD). The alveolar organ with fluorescence and bar code pedigree tracking is constructed and can track the fate transformation of alveolar type II cells (AT2) in a disease model, and the cell pedigree is further decoded, so that a new research basis is provided for early diagnosis and treatment of COPD (chronic obstructive pulmonary disease).

The invention discloses a chronic obstructive pulmonary disease mouse model induced by long-term continuous cigarette smoke exposure as well as a construction method and application thereof, and the construction method comprises the following steps: introducing cigarette smoke with constant concentration into a smoke exposure device, so that a mouse is in a cigarette smoke exposure environment for a long time and maintains normal diet and drinking; performing macroscopic characterization and microcosmic index evaluation on the mouse model with the chronic obstructive pulmonary disease until clinical symptoms of the chronic obstructive pulmonary disease appear; wherein the smoke exposure device comprises a smoke mixing box, a sealed feeding exposure box and a control box, the sealed feeding exposure box provides a cigarette smoke exposure environment, and the control box maintains the stability of the cigarette smoke exposure environment. According to the method, the occurrence cause of the chronic obstructive pulmonary disease of the mouse is simulated by optimizing exposure parameter conditions, the clinical symptoms of the chronic obstructive pulmonary disease of the mouse model are evaluated in multiple dimensions, and the method accords with the symptoms and signs of the chronic obstructive pulmonary disease, is high in similarity and good in reliability, and can be used as a disease model for research of the chronic obstructive pulmonary disease.

The invention provides the method for constructing the secondary glioblastoma organ, so that the secondary glioblastoma organ is successfully constructed, and the defect that the secondary glioblastoma organ cannot be cultured by the current brain glioblastoma organ model is overcome; a research tool closer to the original state of glioma is provided for research on secondary glioblastoma of human brain, and a more reliable research tool is provided for tumor microenvironment research, tumor generation mechanism research, drug effect screening, development and application of disease models and the like of secondary glioblastoma.

The invention discloses a reverse transcriptase, wherein a coding sequence of the reverse transcriptase is derived from a Rattus norvegicus genome. The protein sequence of the protein comprises a protein sequence of a wild type Rattus norveicus, or a protein sequence which is subjected to engineering modification on the basis of the wild type Rattus norveicus, or a protein sequence which is subjected to engineering modification on the basis of the wild type Rattus norveicus. Furthermore, the protein sequence of the gene is as shown in any one of SEQ No.1-55. In a rice genome site and a human HEK 293T cell genome site, the system shows efficient editing efficiency of a guide editor. The invention provides a series of efficient reverse transcriptase components which can be carried on a guide editing system, important bottom-layer technical support is provided for research of point mutation in a genome, creation of a special site disease model and correction of genetic mutation sites, and the reverse transcriptase component has a wide application prospect in the field of precision medicine and precision agriculture breeding.

The invention belongs to the field of biosensing and fluorescence imaging, and particularly relates to a fluorescent probe compound and application thereof. The probe compound is a compound as shown in a formula I or a compound as shown in a formula II, or a pharmaceutically acceptable salt thereof; the structures of the compound in the formula I and the compound in the formula II are shown in the specification, and the definition of each group is shown in any embodiment of the invention. The fluorescent probe compound shows rapid and high reactivity to oxidizing free radicals, the generated fluorescent compound is high in fluorescence intensity and good in stability, and early diagnosis can be carried out on ferroptosis-related disease models such as oxygen-glucose deprivation / reperfusion and drug liver injury on living cell or living body levels and the like. I II

The present invention relates to a biomarker composition for diagnosing stroke or post-stroke neurological damage, for diagnosing the severity of stroke or post-stroke neurological damage, and for predicting stroke prognosis, and the like. Using an ischemic stroke mouse disease model, a tissue damage analysis and a behavioral analysis were performed, and the mRNA level expression was confirmed, and as a result, Prox1 or Dcx could be selected as a biomarker. In addition, it was confirmed that the expression of Prox1 or Dcx increased as the severity of stroke or post-stroke neurological damage was higher, and thus the present invention is expected to be useful for diagnosing stroke or post-stroke neurological damage, for diagnosing the severity of stroke or post-stroke neurological damage, and for predicting stroke prognosis.

This invention relates to a method for constructing a mouse orthotopic model of highly aggregated glioblastoma, belonging to the fields of biomedicine and experimental animal model technology. Glioblastoma cells are resuspended in a composite carrier of a specific ratio of matrix gel and serum-free culture medium, and a low-temperature injection technique is used. Utilizing the thermosensitivity of matrix gel in the mouse intracranial environment (i.e., its phase transition from liquid to gel at a certain temperature), precise colonization and physical locking of tumor cells at target coordinate points are achieved. This method improves upon existing glioblastoma mouse orthotopic model construction techniques that suffer from tumor cell diffusion and loss, low tumor formation rate, irregular tumor morphology, large intragroup variability, and needle reflux due to low viscosity of the inoculation carrier. It provides a good disease model for subsequent screening or evaluation of anti-glioblastoma drugs.

The invention belongs to the technical field of disease model construction, and particularly relates to a construction method of a chronic myelogenous leukemia mouse model with conditional human EZH2 overexpression and RUNX1 knockout. According to the invention, a chronic myelogenous leukemia mouse transgenic mouse model with conditional human EZH2 overexpression and RUNX1 knockout is successfully constructed, the model is induced to be converted from a chronic stage to a sudden change stage, and particularly, the model is a transgenic mouse model which is positive in Lyz2-CreERT2 / EZH2 / RUNX1 and carries BCR-ABL and SCL-tTA. It is proved that a human EZH2 conditional overexpression and RUNX1 knockout chronic myelogenous leukemia mouse transgenic mouse model has feasibility and importance for research on conversion from CML CP to BC samples, and a molecular mechanism for conversion from chronic myelogenous leukemia to a sudden change stage is revealed for research. And a new animal model and a new research idea are provided for understanding of disease progression and development of a new treatment strategy.

A method for the selection of processing parameters for nanomaterial compositions is performed by preparing a baseline model wherein a nanomaterial composition interacts with a disease model. It is then acquired a baseline interaction between the nanomaterial composition and the disease model. A perturbation is applied iteratively by changing at each iteration at least one parameter of the perturbation and acquiring at each iteration a perturbation parameter describing a protein corona formation on said first nanomaterial composition under the effect of the perturbation. Finally, it is identified and selected among the perturbation parameters at least one processing parameter minimizing the formation of protein corona.

The invention discloses a method for constructing astrocytes serving as a smoke disease model, and belongs to the technical field of crossing of stem cells and neuroscience. The method comprises the following steps: S1) reprogramming CD34 + cells in in-vitro PBMCs (peripheral blood mononuclear cells) of smoke disease patients carrying and not carrying RNF213p.R4810K mutation to obtain induced pluripotent stem cells; and S2) directionally inducing and differentiating the induced pluripotent stem cells into astrocytes through a neural progenitor cell way, wherein the obtained astrocytes are the astrocytes capable of being used as the smoke disease model. The astrocyte model prepared by the invention can be used for researching pathogenesis, nerve-blood vessel interaction process and blood-brain barrier (BBB) related functions of smoke diseases, and can be further applied to molecular typing of diseases and in-vitro function evaluation of candidate drugs.

The invention belongs to the technical field of gene editing and disease model construction, and particularly relates to a construction method and application of a Wilson disease animal model. Aiming at a large fragment deletion mutation type which exists clinically in Wilson disease but lacks a corresponding animal model, a CRISPR / Cas9 gene editing technology is utilized, a pair of sgRNAs is specifically designed, an eighth exon region of a mouse Atp7b gene is precisely cut and deleted, and a WD mouse model with deletion mutation (c.2333340delGACGGTGG) of eight basic groups of the eighth exon of the Atp7b gene is successfully constructed. The method can be used for research on pathogenesis of WD, screening of novel diagnostic markers and evaluation of curative effect of therapeutic drugs, is particularly suitable for preclinical evaluation of gene therapy strategies and development of adjuvant therapeutic drugs of targeted NLRP3 inflammasomes, and has great scientific research value and clinical transformation prospect.

The application provides a reprogramming vector for blood cells and application thereof. Through the technical scheme in the disclosure, a low-cost and high-benefit solution can be provided for efficient reprogramming and application of iPSCs, and the wide application of iPSCs in clinical and scientific research fields is significantly promoted, and the iPSCs have important application value and development potential in research and practice in aspects such as disease models, drug screening, cell therapy and the like.

The invention relates to the technical field of organoids, and particularly discloses a chick intestinal tract crypt stem cell separation medium and a separation method and application thereof, and the technical key points are as follows: the separation medium provided by the scheme of the invention can efficiently separate chick intestinal tract crypts with complete structure, good activity and high purity under a low-temperature condition by optimizing component combination; according to the separation method provided by the scheme of the invention, the separation efficiency and the culture quality of the intestinal stem cells and the intestinal organs are remarkably improved through the matching of the separation liquid and the washing liquid. By utilizing the method disclosed by the invention, a high-quality stem cell source can be provided for culture of intestinal organs, drug screening and disease model construction, and the method has important scientific research and application values.

The present invention provides methods of differentiating pericytes from pluripotent stem cells comprising culturing steps in chemically defined culture medium. A population of exogenously derived pericytes from PSCs are also contemplated. Further uses of the exogenously cultured pericytes for an in vitro disease model or in vitro angiogenesis assay are contemplated, including an in vitro 3D model of vasculature.

The invention relates to the technical field of biomedicine, in particular to an in-situ PDX model construction method of peritoneal pseudomucus tumor. According to the in-situ PDX model construction method of the peritoneal pseudomucus tumor, a disease model can be successfully constructed, and the tumor characteristics of the peritoneal pseudomucus tumor are highly restored. Particularly, the model can continuously form a large amount of mucus in the abdominal pelvic cavity and accurately reproduce the distribution and expression characteristics of the peritoneal false mucus tumor in the abdominal pelvic cavity, so that a reliable experimental basis is provided for exploring a treatment strategy of the peritoneal false mucus tumor.

The invention relates to the technical field of immune research, in particular to a disease model macrophage in-vivo programming method which comprises the following steps: constructing a target transcription factor plasmid containing a peptide barcode protein sequence; wherein the target transcription factor plasmid contains the peptide barcode protein sequence and a tdTomato fluorescent protein sequence; forming a target transcription factor mRNA in vitro by taking the target transcription factor plasmid as a template; combining and embedding the target transcription factor mRNA into lipid nanoparticles; and injecting the lipid nanoparticles into the body of a mouse through intravenous injection, enabling the lipid nanoparticles to be stably expressed in the body of the mouse, taking the lipid nanoparticles as a disease animal model, and collecting a tissue sample. According to the invention, specific transcription factors and a digital coding technology are utilized, so that the efficiency of transforming the B cells into the macrophages is improved. In-vivo imaging and evaluation of transcription factors are realized through in-vivo delivery of lipid nanoparticles. And a new technical means is provided for process monitoring and mechanism research of a disease model.

The invention discloses an intelligent cerebral apoplexy cell state recognition method based on Cellpose and fluorescence image analysis and application of the intelligent cerebral apoplexy cell state recognition method in drug effect screening. The intelligent cerebral apoplexy cell state recognition method comprises the following steps: S1, establishing a cerebral apoplexy ischemia reperfusion cell model; s2, adding a nitrite fluorescent probe into the cells subjected to ischemia reperfusion treatment to obtain a fluorescent microscopic image of the cells; s3, importing the fluorescence microscopic image into a Cellpose model for cell segmentation, and obtaining an ROI region of each cell; s4, extracting fluorescence intensity characteristics and cell morphological characteristics in the ROI region of each cell; and S5, inputting the extracted multi-dimensional features into a classification model for cell state recognition, so as to distinguish a single cell into three states of health, mild injury or serious injury. According to the method, a complete link of image acquisition-Cellpose analysis-feature extraction-state discrimination-drug effect evaluation is constructed, and the practical application value of a deep learning image algorithm in disease model research and drug screening scenes is expanded.

The invention discloses a novel ALS disease model and application thereof in screening ALS treatment drugs, and belongs to the technical field of disease models and drug screening. According to the method, induced pluripotent stem cells derived from a sporadic ALS patient are directionally induced into spinal cord organs, the spinal cord organs and immune cells derived from the same donor form a co-culture system, the co-culture system is infected with viruses, and the ALS disease model is successfully constructed. According to the method, ALS diseases are simulated on the three-dimensional level, ALS disease models of all stages (including the initial stage of the diseases) are obtained, and the pathological features of all the disease stages of ALS (especially 90% or above of sporadic ALS) can be accurately reflected; the problem that key disease-promoting factors in the early stage of the disease cannot be obtained due to complete disorder of downstream signal channels in the end stage of the disease after definite diagnosis caused by slow definite diagnosis of hidden disease onset of the ALS is solved.