109 results about "In vitro model" 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 biological medicines, and particularly discloses a novel application of Epiblastatin A in preparation of a medicine for relieving cisplatin-induced acute kidney injury. The application disclosed by the invention for the first time reveals that by inhibiting the activity of Casein Kinase 1 (CK1) kinase, Epiblastine A can obviously relieve renal dysfunction and kidney tissue structure damage caused by cis-platinum. Experiments prove that the compound can effectively reduce serum creatinine and urea nitrogen levels, significantly improve renal tubule pathological injury, down-regulate expression of renal injury early-stage specific biomarkers KIM-1 and NGAL, and definitely inhibit renal tubule epithelial cell apoptosis in an in-vitro model. The invention also provides a specific pharmaceutical composition containing the Epiblastin A and a preparation form of the specific pharmaceutical composition. The invention provides a brand new mechanism and a high-efficiency and low-toxicity treatment strategy for clinically preventing and treating cisplatin dosage-limited renal toxicity, and has important clinical application value.

The invention discloses a combined evaluation method for in-vitro and in-vivo red fading and relieving effects of cosmetic raw materials, which comprises the following steps: in-vitro model evaluation: inducing a human immortalized umbilical vein endothelial cell model by adopting TNF-alpha, and detecting the secretion amount of a prostacyclin I2 factor to obtain a score of an in-vitro model; in-vivo model evaluation: adopting a chick embryo chorioallantoic membrane model, and obtaining a score of the in-vivo model according to the degree of vasoconstriction caused by the to-be-detected raw material sample acting on the chick embryo chorioallantoic membrane; and comprehensive evaluation: based on the score of the in-vitro model and the score of the in-vivo model, comprehensively evaluating the red fading and relieving efficacy of the raw material sample to be tested. According to the technical scheme, the defect that in the prior art, no cosmetic raw material soothing efficacy evaluation method for the vascular reactivity increasing mechanism exists is overcome, efficacy evaluation is more accurate, the method has the advantages of being comprehensive, short in test period, easy to operate, economical and convenient, and an effective reference basis is provided for screening and compounding schemes of red fading and soothing raw materials.

The invention discloses a three-dimensional heart chip device. The device comprises a pneumatic control layer, a flexible functional film layer and a microfluidic culture layer, wherein the flexible functional film layer is integrated with a stretchable microelectrode array with a snakelike structure, and the microfluidic culture layer is arranged on the flexible functional film layer; wherein the pneumatic control layer is used for driving the flexible functional film layer to generate periodic deformation and applying mechanical tensile strain to cardiac muscle tissues in the microfluidic culture layer, and meanwhile, the stretchable microelectrode array of the snake-shaped structure synchronously deforms along with the flexible functional film layer and keeps conformal electric contact with the cardiac muscle tissues, so that the mechanical tensile strain is applied to the cardiac muscle tissues in the microfluidic culture layer. The electrophysiological signals are monitored in the dynamic mechanical stretching process. The technical problem that in the prior art, stable electrophysiological monitoring cannot be carried out while dynamic mechanical stretching is applied is solved, physiologically-related mechanical stimulation can be applied to three-dimensional cardiac muscle tissue to promote maturity of the three-dimensional cardiac muscle tissue, and long-term and stable in-situ electrophysiological signal monitoring is achieved in the process; and a more accurate in-vitro model is provided for drug screening and disease research.

The application belongs to the technical field of bone organ preparation, and relates to a bone organ for promoting nerve growth and a construction method and application thereof. The construction method comprises the following steps: S1, immersing a hydrogel mineralization scaffold with conductive polymer induction in an EDC / NHS crosslinking agent, and sterilizing and crosslinking; S2, immersing the crosslinked product in a culture medium and culturing, inoculating bone organ seed cells, and replacing the culture medium with a complete culture medium for adaptive culture; and S3, transferring to an osteogenic induction culture medium for osteogenic induction, so as to obtain the bone organ for promoting nerve growth. Compared with the prior art, the bone organ constructed by the application has good biological activity, osteogenic activity and bone conduction, can effectively promote the growth of peripheral nerves, provides a reliable in-vitro model for bone-nerve interaction research, and provides a new technical path and theoretical basis for bone-nerve integrated regeneration and repair.

Cannabidiol, or a pharmaceutically acceptable prodrug, derivative, salt, or solvate thereof, for use in treating or preventing recurrent pericarditis is disclosed. The effectiveness of cannabidiol in attenuating the levels of interleukin-1β (IL-1β) and IL-6, and the transcription of pro-IL-1β and NLRP3 mRNA in in vitro models are disclosed. Also disclosed are experiments showing cannabidiol to be effective in preventing an increase in the pericardial space (which is an indicator of pericardial effusion) and the pericardial thickness in an in vivo mouse model of pericarditis.

The application discloses a human vagina micro-ecological in-vitro model and a construction method and application thereof, and relates to the technical field of medicines.The application discloses a human vagina micro-ecological in-vitro model and a construction method and application thereof.Firstly, a mixed cell group of the vaginal epithelium containing a complete cell lineage and capable of long-term subculture is successfully established, a technical bottleneck that primary vaginal epithelial cells are difficult to expand is solved, and ideal seed cells are provided for model construction.Secondly, a highly-bionic vaginal mucosa model having a tight connection structure, rich layers and an orderly arrangement is constructed, and the physiological barrier and immune regulation functions of the vaginal mucosa are truly simulated.Finally, by introducing lactobacillus symbiosis and confirming the response to estrogen, a multi-element in-vitro research model integrating the anatomical structure of the vagina, resident flora, local immunity and endocrine regulation is constructed, and a reliable technical platform is provided for in-depth exploration of the mechanism of vaginal micro-ecological disorder and efficient screening of related drugs.

The invention belongs to the technical field of nerve cell culture, and particularly relates to application of a culture medium in co-culture of SVGP12 and HMEC-1. The culture medium is based on a mixture of DMEM and MCDB 131 according to a ratio of 1: 1, HEPES, Glutamax, N2, G-5, B-27, hEGF, penicillin-streptomycin and fetal calf serum are added, and the components are prepared according to a specific final concentration. When being applied to a Transwell co-culture model, the culture medium can meet the nutritional requirements of two cells at the same time, maintain the normal growth and form of the cells, and promote the transmission of functional signal molecules. Experiments show that the culture medium can significantly up-regulate expression of ABCG2, CYP2D6, CYP1B1 and CYP2E1 in HMEC-1 (P is less than 0.05), provides a reliable in-vitro model for researching an interaction mechanism of two cells, and assists drug screening and disease simulation.

The invention belongs to the technical field of biological medicine, and discloses a toxoplasma gondii in-vitro merozoite induction model and a construction method thereof. The invention discloses application of a glutamine antagonist to induction of differentiation of toxoplasma gondii tachyzoite to merozoite or preparation of a product for induction of differentiation of toxoplasma gondii tachyzoite to merozoite, and finds that the glutamine antagonist JHU083 can induce differentiation of toxoplasma gondii virulent strain RH and attenuated strain ME49 tachyzoite into merozoite under in-vitro conditions for the first time. A reliable in-vitro model is provided for researching the key initial stage of sexual reproduction of the toxoplasma gondii, and an irreplaceable research foundation is laid for the key link of targeted blocking of parasite transmission.

The invention belongs to the field of biomedical engineering, and particularly relates to an amino acid modified molybdenum disulfide artificial extracellular matrix and application. Preparing an amino acid modified molybdenum disulfide artificial extracellular matrix, inoculating the neuron in-vitro model PC12 cell suspension onto the artificial extracellular matrix, and constructing the nerve cell in-vitro differentiation model. The artificial extracellular matrix with good biocompatibility is obtained by constructing different kinds of amino acid modified molybdenum disulfide nano material artificial extracellular matrixes as the artificial extracellular matrix of nerve cells PC12, and the artificial extracellular matrix improves the activity of the PC12 cells and promotes the differentiation of nerve cell synapses; a neuron in-vitro model with high differentiation and higher activity is constructed, and the artificial extracellular matrix can be used for nerve regeneration and neural engineering.

The invention discloses a method and a device for detecting the energy metabolism level of cells and organoids. According to the detection method disclosed by the invention, the acidification rate and the oxygen consumption rate of the extracellular microenvironment are obtained through signal response of hydrogen ions and the dissolved oxygen sensing electrode, and meanwhile, a dynamic detection microcavity is constructed by utilizing a metabolism detection device, so that the sensitivity and the accuracy of measurement are improved. By adopting different drug intervention schemes, the energy metabolism pathway of the cells can be regulated and controlled, so that the evaluation of the two energy metabolism pathways of glycolysis and aerobic respiration of the cells is realized. The in-situ and high-sensitivity cell energy metabolism detection method and device are provided by detecting the concentration change of the specific analyte of the extracellular microenvironment, the method has the universality of two-dimensional and three-dimensional cell models, a brand new platform is provided for scientific research and drug effect screening based on an in-vitro model, and the application prospect is wide.

The application provides a method for constructing an immortalized bovine nasopharyngeal tonsil epithelial cell line and application, and belongs to the technical field of cell engineering. The healthy yellow bovine nasopharyngeal tonsil is used as a material, and the primary epithelial cells are separated by enzyme digestion. The 3rd to 4th generation cells are taken, infected with SV40 T lentivirus (MOI=80), and screened by 1.5 μg / mL puromycin to obtain a stable immortalized cell line. The cell line is continuously passaged to the 50th generation and still stably expresses SV40LT protein and epithelial specific markers CD326, CD324 and KRT8, maintains a typical pavement stone-like morphology, normal proliferation and no malignant transformation. Infection experiments show that the cell is highly sensitive to FMDV, supports efficient replication, and the virus titer can reach 10 7.5 TCID 50 / mL. The application overcomes the defects of the existing immortalized bovine nasopharyngeal tonsil epithelial cell model, such as limited source, large batch difference, low sensitivity and poor persistence, and provides a stable and reliable in vitro model for the research of foot-and-mouth disease prevention and control related mechanisms, drug screening and vaccine development.

The invention provides a construction method of an HBV (Hepatitis B Virus) infection model based on multilineage hepatobiliary organs, which comprises the following steps: differentiating human pluripotent stem cells to form immune cell-containing hepatobiliary organs for further maturation culture, and enabling the hepatobiliary organs to be mature in function and highly express NTCP (Natriuretic Therapeutic Cells) by using a maturation culture medium containing bile metabolism substrate cholesterol and deoxybear bile acid, so as to obtain the HBV infection model based on the multilineage hepatobiliary organs. And then infecting the mature organs by adopting an HBV virus strain generated by HepAD38 cells to construct an in-vitro model which is susceptible to HBV and can maintain an infection state for a long time. According to the construction method, the liver infection and immune interaction microenvironment can be simulated more truly, and a stable and reliable experimental platform is provided for HBV infection mechanism research and anti-HBV drug screening.

The Cardio-Tox Tissue Engineered Model (TEEM) invention provides a robust in vitro model for cardiotoxicity evaluation using three-dimensional (3D) human heart microtissues to quantify dose-dependent changes in electromechanical activity, resulting in a comprehensive cardiotoxicity and arrhythmia risk assessment of test compounds. The invention also provides a predictive in vitro screening platform for pro-arrhythmic toxicity testing using human three-dimensional cardiac microtissues. The invention enables the screening of environmental and pharmaceutical compounds, chemicals, and toxicants to establish safe human exposure levels.

This invention belongs to the field of biomedical technology, specifically relating to a method and application for preparing CAR-T cells with enhanced persistence based on the piggyBac transposon system combined with electroporation. This invention, through specific piggyBac dual plasmid ratios and electroporation conditions, can improve transfection efficiency to over 80%, and the integration site is more likely to be located in a safe genomic region, supporting long-term stable CAR expression. Furthermore, cell expansion kinetics are significantly improved, with expansion exceeding 120-fold within 15 days. The results of the examples show that CAR-T cells prepared using the method described in this invention exhibit significantly enhanced tumor cell killing ability and cytokine secretion levels in in vitro models, and are rich in memory T cell subsets (Tcm / Tem) with greater persistence and self-renewal potential. This indicates that this invention not only changes the method of gene delivery but also shapes the intrinsic properties of cells, enabling them to acquire potentially superior in vivo persistence and long-term anti-tumor capabilities.

The present invention relates to an in-vitro model of thehuman gut microbiome, the model comprising a culture of the gut microbiome, wherein the model has a cumulative enzymatic coverage of more than 85% of the gut microbiome of a healthy human. The model facilitates metabolic modeling and enables a better understanding of the structure and function of the human gut microbiome as well as of modifications of xenobiotics by intrinsic gut microbiota, such as biotransformation and bioaccumulation. It can further be used to study the effects of variations in nutritional conditions. Importantly, the invention can also be used for diagnosing a disease, such as a gastrointestinal disorder,a proliferative disease, a metabolic disorder, a cardiovascular disease, an immunological disease, and an infectious disease.

The invention discloses a pig jejunum crypt separation and organoid model construction method, and belongs to the technical field of animal tissue separation and culture. The method specifically comprises the following steps: intestinal tract collection, cleaning solution preparation, intestinal tract treatment, tissue trimming, tissue cleaning, digestive juice preparation, tissue digestion, cell mass separation, crypt separation, crypt counting, crypt collection, crypt inoculation and organoid culture. The porcine jejunum organ obtained by the method has a typical three-dimensional cystic structure, has complete epithelial polar and functional cell types, including microvillus, goblet cells, panning cells and endocrine cells, and shows good barrier function and drug metabolic activity. A reliable in-vitro model platform is provided for pig intestinal development, nutrient absorption, pathogen infection mechanism research and precision medical treatment, meanwhile, a technical foundation is laid for large-scale application of organoids, and the method has important scientific significance and clinical value.

The application relates to a method for separating and primary culturing of monopterus albus liver cells. The method comprises the following steps: obtaining monopterus albus liver tissue, perfusing the liver tissue with HBSS buffer solution without calcium and magnesium until blood is completely drained; removing the capsule, blood clots and connective tissue after rinsing, and cutting the liver tissue; sequentially using 0.02% EDTA solution, 0.25% trypsin solution and 50 U / mL collagenase IV working solution to perform three-step digestion treatment on the liver tissue fragments; obtaining a cell suspension by blowing and filtering after the digestion is completed, and then performing gradient centrifugal purification of the cells at rotating speeds of 600 rpm, 250 rpm and 80 rpm in sequence; and finally resuspending the cells in DMEM / F12 culture medium containing 10% FBS and inoculating and culturing the cells. The monopterus albus primary liver cells obtained by the method have the advantages of large quantity, high activity and high purity, and provide a reliable in-vitro model for monopterus albus related research.

The invention provides a novel human distal bile duct cancer cell line with TP53 missense mutation and application, the novel human distal bile duct cancer cell line CBC3T-3 is established, the cell line is preserved in the China Center for Type Culture Collection (the preservation number is CCTCC NO: C202555), the uniqueness and stability of the cell line are proved through STR typing and karyotype analysis, and the TP53 missense mutation novel human distal bile duct cancer cell line has the advantages that the TP53 missense mutation novel human distal bile duct cancer cell line CBC3T-3 can be used for preparing the TP53 missense mutation novel human distal bile duct cancer cell line CBC3T-3; and a plurality of driver gene mutations including TP53 missense mutation are carried. The CBC3T-3 has strong proliferation, invasion and migration capabilities, has high tumor formation rate in immunodeficient mice, is resistant to cis-platinum and sensitive to paclitaxel and gemcitabine, and provides an experimental basis for selection of clinical chemotherapy regimens. According to the model, the TP53 missense mutation type distal bile duct cancer in-vitro model is successfully established, and a key experimental platform is provided for deeply researching the drug resistance mechanism of the TP53 missense mutation type distal bile duct cancer and developing an individualized treatment strategy aiming at the subtype of the TP53 missense mutation type distal bile duct cancer.

The present invention relates to a method for producing an in vitro model of a human neurological disease. Furthermore, the invention relates to the in vitro model produced according to the method disclosed herein. The invention also relates to the use of the in vitro model in drug testing. In addition, the invention relates to methods for testing the efficacy of drugs in preventing, delaying, and alleviating a human neurological disease.

The invention discloses a bronchial asthma treatment drug based on RBX1 inhibition and application thereof, and finds that expression of RBX1 in asthma patients and disease models is up-regulated, and the expression level of RBX1 is in positive correlation with airway epithelial barrier injury and active oxygen accumulation degree, which indicates that RBX1 is a key new target for asthma treatment. On the basis of the compound Z226407860 and the compound Z4189742053, the condition that the expression of the RBX1 can be effectively inhibited is further proved. In an in-vitro model, the compounds not only significantly reduce the protein level of RBX1, but also show a good synergistic treatment effect: the compounds can effectively relieve the injury of the airway epithelial barrier and promote the integrity recovery of the barrier, and can inhibit the excessive accumulation of active oxygen at the same time, so that the asthma process is intervened from two correlative core pathological links. The research provides a solid experimental basis and a candidate compound with a clear prospect for developing a therapeutic drug which aims at bronchial asthma and has a brand new action mechanism.

The present invention introduces an approach to neural tissue engineering by developing vascularized neurospheres (VNSs) through novel methods, systems, and kits. These VNSs are generated using a unique blend of neural, vascular, and supportive cells, cultured in specialized hydrogel-containing media. The VNSs feature an integrated vascular network, serving as a physiologically relevant in vitro model. The invention also offers a grafting technique for therapeutic applications in neural disorders and injuries, as well as facilitating brain-computer interfacing. Beyond therapeutics, the VNSs have broad applications in drug development, synthetic biology, disease modeling, and more. The invention also includes kits for VNS generation and methods for interfacing VNSs with probes for applications of brain-computer interfacing. Overall, this patent represents a significant leap in the field, bridging the gap between in vitro and in vivo systems and opening new avenues in research, therapeutics, and bioengineering.

The invention belongs to the technical field of functional nutritional composition research, and relates to a nutritional composition and application thereof, in particular to a nutritional composition and non-therapeutic application thereof in retarding brain aging injury. The nutritional composition comprises lactoferrin and 3 '-sialyllactose as essential active ingredients. Experiments find that in an in-vitro model, combined treatment of lactoferrin and 3 '-SL can relieve oxidative stress aging damage of Neuro-2a nerve cells induced by H2O2, and the oxidative stress aging damage is mainly manifested as follows: the content of an oxidative stress marker MDA is decreased, the SOD activity is increased, brain aging-related secretory phenotype proinflammatory factors IL-6 and TNF-alpha are decreased, and an anti-inflammatory factor IL-10 is increased. Oxidative stress is considered to be one of the important mechanisms of brain aging and neurodegenerative diseases. The invention finds that the nutritional combination of lactoferrin and 3 '-SL slows down brain aging injury in the aspects of regulating oxidative stress and inhibiting neuroinflammation.

This invention provides a choline-inspired amphiphilic polymer nanodisc, the raw material for which includes a polymethacrylate copolymer; the polymethacrylate copolymer is prepared by polymerization of 2-methacryloyloxyethylphosphorylcholine and hexyl methacrylate. This choline-inspired amphiphilic polymer nanodisc possesses efficient BBB-crossing and brain-targeting capabilities. It can efficiently cross the BBB via choline transport receptors in both in vitro BBB models and in vivo in mice. The disc-shaped morphology and nanoscale size of the choline-inspired amphiphilic polymer nanodisc promote effective penetration of blood vessels, matrix, and cell barriers, exhibiting excellent solid tumor infiltration capabilities. This invention also provides a method for preparing the above-mentioned choline-inspired amphiphilic polymer nanodisc and its application in the preparation of brain tumor detection reagents or drugs for the prevention and treatment of brain tumors.

The application discloses application of recombinant human USP13 protein in preparation of a drug for treating diabetic cardiomyopathy, and belongs to the technical field of biological medicines.The application first proves that the expression of USP13 in myocardial tissue and serum is significantly reduced under the condition of diabetic cardiomyopathy (DCM); the recombinant human USP13 protein can obviously reduce the level of myocardial cell active oxygen, reduce the expression of NLRP3 inflammasome and active Caspase3 protein in a DCM in-vitro model induced by high sugar / palmitic acid; in a DCM mouse model injected with the recombinant human USP13 protein through a tail vein, myocardial cell apoptosis is inhibited, and the expression of NLRP3 inflammasome and the synthesis of TNF-alpha in the heart of the animal are reduced.USP13 is a human protein, has no obvious toxic side effects, is high in safety, is small in immunological rejection in theory, provides a new effective drug for the clinical treatment of DCM, and has important clinical application value and industrialization prospect.

The Cardiac Tissue Engineered Model (TEEM) invention provides a robust in vitro model for cardiotoxicity evaluation using three-dimensional (3D) human heart microtissues to quantify dose-dependent changes in electromechanical activity, resulting in a comprehensive cardiotoxicity and arrhythmia risk assessment of test compounds. The invention also provides a predictive in vitro screening platform for pro-arrhythmic toxicity testing using human three-dimensional cardiac microtissues. The invention enables the screening of environmental and pharmaceutical compounds, chemicals, and toxicants to establish safe human exposure levels.

The application belongs to the field of organoid culture, and specifically discloses a culture method of monkey gastric organoids, and discloses a culture medium for culturing monkey gastric organoids, wherein the culture medium comprises (1) an expansion culture medium and (2) a complete culture medium; the application screens key regulation factors and appropriate concentrations for the growth of monkey gastric organoids according to the biological characteristics and growth characteristics of monkey gastric organs, optimizes the culture method, maintains the activity of monkey gastric organoids in vitro for a long time, and can be stably passaged, frozen, recovered, and cultured on a large scale for functional experiments, so as to provide a primate in-vitro model for the research on the mechanism of gastric diseases, toxicity testing, drug research and development, and the like.

The invention discloses an integrated bone marrow microenvironment-bone tumor organoid micro-fluidic chip, and relates to the technical field of bone tumor in-vitro research models, the key points of the technical scheme are as follows: the chip is of a double-layer structure, is separated by a porous membrane in the middle, and is matched with a micro-fluidic structure formed by combining a plurality of cavities; the multiple chambers comprise a tumor organ chamber, a blood vessel communication chamber and a marrow chamber; the bone marrow upper-layer chamber and the bone marrow lower-layer chamber are separated by a porous membrane, the bone marrow upper-layer chamber is a nutrition supply layer, the bone marrow lower-layer chamber is a cell culture layer, and the porous membrane allows gradient diffusion of micromolecule nutrition / cell factors, so that nutrition supply of the bone marrow upper-layer chamber to the bone marrow lower-layer chamber is realized. The chip can reproduce a complex three-dimensional structure of the BMME, realizes dynamic microenvironment control and real-time visual monitoring, and provides a more accurate in-vitro model for research of bone tumor organs.