168 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 discloses a bone organ chip loaded with a microelectrode array and a use method, and relates to the technical field of organ chips, the bone organ chip is made of a polydimethylsiloxane material, and the bone organ chip comprises a three-layer structure: a top layer, a middle layer and a bottom layer; the middle layer comprises two blood vessel micro-channels, two osteoblast micro-channels and a neuron micro-channel, the channels are separated through columns and micro-columns, and cell interaction in a bone micro-environment is simulated; a microelectrode array is arranged on the top layer and is used for monitoring electrical activity of neurons in real time; the bottom layer is provided with a pneumatic valve which can control opening and closing of the micro-channel. According to the invention, nerve and blood vessel channels are introduced, a nerve-blood vessel-immunoregulation bone microenvironment is constructed, physiological processes such as osteogenesis-blood vessel formation coupling and the like are embodied, intervals of different sizes can enhance intercellular crosstalk and allow blood vessels and neuron axons to selectively pass through, so that the chip can simulate a complex microenvironment of bone tissues, and the microenvironment of the bone tissues can be simulated; and an efficient and accurate in-vitro model is provided for bone disease mechanism research and drug development.

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 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 provides a skin fibrosis in-vitro model as well as a construction method and application thereof, and relates to the technical field of cell biology. According to the skin fibrosis in-vitro model construction method, a full-thickness skin model with an epidermal layer and a corium layer is constructed by utilizing a cell culture chamber or a cell culture micro-fluidic chip, and the skin fibrosis in-vitro model is obtained by performing induced culture through a chemical reagent, a growth factor or mechanical stress while the corium layer is constructed. According to the construction method, the problem that an existing in-vitro model can only reflect early-stage fibrosis skin characteristics and cannot effectively simulate fibrosis skin formed under the condition of a long disease course is solved. Besides, the corium layer and the epidermal layer in the full-thickness skin structure are not separated, isolated and differentiated through a porous membrane, corium layer fibrosis can generate more direct influence on the epidermis, richer inflammatory factor expression change of the whole model is obvious, and the simulation degree of skin fibrosis diseases is higher.

The invention belongs to the technical field of organ chips, provides an in-vitro brain lymphatic system chip construction method and a brain lymphatic system chip, and makes up the blank of brain lymphatic system in-vitro model construction. The method comprises the following steps: firstly, carrying out flushing treatment, disinfection and sterilization treatment, rinsing treatment and cell culture on a micro-fluidic channel in the chip; then, in the treated micro-fluidic channel, blood vessel channel cell inoculation and brain parenchyma channel cell inoculation are carried out; and finally, in the blood vessel channel, perfusion is carried out once at a preset flow speed every preset duration, shear force stimulation simulating pulse type blood flow is provided, basic formation of in-vitro cerebral vessels is achieved after continuous perfusion, perfusion is carried out once at a preset flow speed every preset duration, shear force stimulation simulating pulse type blood flow is provided, and in-vitro cerebral vessels are formed. Intermittent liquid flow simulating arterial fluctuation is achieved.

The invention discloses a three-dimensional biological printing meniscus model for simulating the mechanical heterogeneity of a natural meniscus and a preparation method of the three-dimensional biological printing meniscus model. The model comprises a structural PCL fiber frame (red region fiber circumferential arrangement and white region radial arrangement, and simulates the trend of natural collagen fiber), regional specific hydrogel (the red region contains GelMA, HAMA and SilMA, and the white region contains GelMA and HAMA), and partitioned cell components (the red region contains BMSC, HUVEC and CTGF, and the white region contains BMSC and TGF-beta3), the PCL frame and the hydrogel cooperate to form a mechanical gradient, and the compression modulus from the red region to the white region is simulated to continuously change. The preparation method comprises the following steps: designing a three-dimensional model, printing a PCL framework, preparing hydrogel and a cell suspension, preparing bio-ink, filling and printing by multiple nozzles, curing and culturing in vitro to obtain a target model. Compared with the prior art, the method has the advantages that the key problem of mechanical mismatch of an existing support is effectively solved, the model better fits a natural meniscus in mechanical property and biological function, and a more adaptive in-vitro model support is provided for individualized meniscus repair and transplantation substitution research.

The invention relates to the technical field of biology, in particular to a composition, a culture medium and application of the culture medium in intestinal organ culture. The glucosamine hydrochloride is applied to culture of intestinal organs for the first time, organ proliferation and differentiation can be effectively promoted, and the number of formed organs is remarkably increased. On the basis, a three-dimensional culture system for staged time sequence induction in a postpartum newborn period, a child period and an adult period is constructed, and accurate construction of intestinal organs of the mice in different development periods is realized by combining dynamic regulation and control of an extracellular matrix and combinatorial optimization of growth factors; and an in-vitro model covering multiple time sequence stages is provided for intestinal development mechanism research and disease modeling.

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.

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 invention is suitable for the technical field of insect genetic engineering, and provides an HparGR22 gene and application thereof in establishing a phlorizin recognition in-vitro model. The DNA sequence of the HparGR22 gene is as shown in SEQ ID No: 1, and the coded amino acid sequence is as shown in SEQ ID No: 2. The gene has the highest expression quantity in the lower labial tassel of a holotrichia parallela larva, an oocyte injected with cRNA of the gene has obvious response to stimulation of phlorizin with different concentrations, it is clear that a product expressed by the HparGR22 gene can recognize phlorizin, and the function of the HparGR22 gene as the phlorizin receptor gene of the holotrichia parallela is determined. In view of the fact that the phlorizin recognition function of the gene is clear for the first time, a basis is provided for application of the gene in establishment of a phlorizin recognition in-vitro model, a foundation is laid for exploring the potential value of the gene in the field of gene engineering for preventing and controlling holotrichia parallela, and meanwhile a new direction is opened up for further research and application of the gene.

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.

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.

To provide a biomimetic system that mimics the human proximal tubule, which is a high-performance in vitro model that can reproduce the physiological functions of the human proximal tubule in vitro and can be used as an evaluation tool in drug discovery.SOLUTION: The present invention provides a proximal tubule biomimetic system comprising a microfluidic device and LTL-positive cells derived from pluripotent stem cells contained within the microfluidic device. The microfluidic device has (A) a device body, (B) a first chamber provided in the device body, (C) a second chamber provided in the device body, and (D) a porous membrane positioned between the first chamber and the second chamber, separating the first chamber from the second chamber. The LTL-positive cells are contained within the first chamber and adhere to a first surface of the porous membrane facing the first chamber. The first surface of the porous membrane is coated with an extracellular matrix.SELECTED DRAWING: None

The invention relates to the technical field of organs, and discloses a culture medium and a method for constructing and culturing vascular organs and application of the culture medium and the method. By utilizing the culture medium set provided by the invention, the vascular organoid with complete functions and structures can be constructed through iPSC induction, and matrigel does not need to be adopted in the construction and culture process, so that the construction operation of the organoid is greatly simplified, and the cost is saved. The blood vessel organoid constructed by the invention has a basic structure of a blood vessel, has corresponding response to medicine components which are known in the field and have influence on the blood vessel, and can be used as a blood vessel in-vitro model.

The present invention provides usage of boric acid in a method of brittle bone disease treatment. In the present invention, moreover, there is a new brittle bone disease (osteogenesis imperfecta) in-vitro model for observing the effect of boric acid on brittle bone disease.

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 an in-vitro model for evaluating the performance of a remineralization-promoting material, a construction method and application thereof, and comprises the following steps: (1) gel preparation; (2) model assembly. The in-vitro model can effectively and accurately evaluate the performance of the remineralization-promoting material, has the characteristics of good evaluation effect, strong distinguishability, strong repeatability, good uniformity, stable and efficient evaluation system, and can complete remineralization of multiple test groups in a short time (5-7 days).

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.

The present invention discloses the use of tazemetostat in the preparation of drugs for treating acute kidney injury, and belongs to the field of biomedicine technology. Specifically, it also relates to the use of tazemetostat in the preparation of drugs for reducing the mRNA expression levels of lipocalin Ngal, kidney injury factor KIM1, chemokine MCP-1, inflammatory factors IL-1β, IL-6, and TNF-α, and the use of tazemetostat in the preparation of drugs for reducing the levels of creatinine and urea nitrogen in serum. Beneficial effect: The present invention proposes a new drug use of tazemetostat in the treatment of acute kidney injury. It effectively reduces the inflammatory response and renal damage level caused in an in vitro model of acute kidney injury; it can also reduce the expression levels of inflammatory factors IL-6 and chemokine MCP-1 in the serum of an in vivo animal model of renal injury, improve the kidney tissue pathology of animals with acute kidney injury, inhibit the protein expression of KIM1 in kidney tissue, reduce the levels of creatinine and urea nitrogen in kidney tissue serum, and has a potential therapeutic effect on acute kidney injury.

The present application relates to the field of biotechnology, and particularly relates to a composition, a culture medium and application thereof in intestinal organoid culture. The present application first applies glucosamine hydrochloride to the culture of intestinal organoids, which can effectively promote organ proliferation and differentiation, and significantly increase the number of organ formation. On this basis, the present application constructs a three-dimensional culture system for postpartum neonatal, childhood and adult stages, and realizes the precise construction of mouse intestinal organoids at different development stages by combining dynamic regulation of extracellular matrix and optimization of growth factor combinations, thereby providing an in-vitro model covering multiple time sequences for intestinal development mechanism research and disease modeling.

The invention discloses a fugu rubripes primary hepatocyte efficient separation and culture method suitable for different growth stages, and belongs to the technical field of hepatocyte culture. Through the optimized pre-cleaning and enzyme digestion steps, the difficulty in hepatocyte separation caused by high-fat liver is effectively overcome, stable acquisition of the fugu rubripes primary hepatocytes at different growth stages is realized, dependence on juvenile fish which is not subjected to feed conversion is not needed, and the applicability of experimental materials is greatly widened. According to the method, the cell yield is high, the growth state is stable, and a reliable in-vitro model is provided for fugu rubripes lipid metabolism research and nutritional immune regulation.