215 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 provides a method for constructing a chronic unpredictable negative stress model, the chronic unpredictable negative stress model and application of the chronic unpredictable negative stress model in mental disease research, and belongs to the field of brain organ in-vitro model construction. Through dual verification of a microelectrode array and single cell sequencing, an in-vitro brain organ model capable of simulating chronic unpredictable negative stress core characteristics is successfully constructed, and a potential neural network function reconstruction mechanism and a cell molecule basis of the in-vitro brain organ model are disclosed. The comprehensive research normal form breaks through the limitation of a traditional single technology platform, and an accurate and efficient innovative research platform highly related to human is provided for pathogenesis research of mental diseases, especially chronic stress related diseases such as depression and anxiety and development of novel treatment strategies.

The invention belongs to the technical field of biomedicine, and discloses an immortalized cell line for human kidney suspected cell carcinoma as well as a culture method and application of the immortalized cell line. The immortalized cell line for the human kidney suspected cell carcinoma, disclosed by the invention, is named as a human kidney suspected cell carcinoma cell line Loya a-710 (Homo sapiens), the classification name of the Latin name of the immortalized cell line is Chromophore sensor cell carcino: Loya a-710, and the preservation number of the immortalized cell line is CCTCC (China Center for Type Culture Collection) NO: C2025123. The human kidney suspected cell carcinoma cell line Loya-710 disclosed by the invention is used as an in-vitro model to rapidly grow in tissue culture, and mitochondrial mutation characteristics, a vesicle structure and a classical immunohistochemical marker of ChRCC are reserved. The invention provides a precious tool for further research on ChRCC heredity, molecules and biological characteristics, and also provides a powerful new model for mitochondrial diseases.

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 invention provides a method for generating platelet-like particles based on MEG-01 cells and evaluating functions of the platelet-like particles, and belongs to the technical field of cell biology. The invention provides a method for generating PLPs by inducing MEG-01 cells. According to the method, the MEG-01 cells are differentiated into PLPs with platelet characteristics by utilizing a PMA inducer. The PLPs disclosed by the invention has characteristic surface marker expression consistent with that of natural platelets, can respond to stimulation of an activator ADP, and shows biological functional characteristics of the platelets. The method for generating PLPs is simple and economical, experimental conditions are highly controllable, and an economical, efficient and repeatable in-vitro model is provided for functional research related to platelets.

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 relates to the technical field of preparation of organoids, and discloses a method for regulating and controlling human induced pluripotent stem cells to differentiate into kidney organoids, which comprises the following steps: resuspending a cell cluster formed by human induced pluripotent stem cells by using a basic differentiation culture solution containing a GSK-3beta inhibitor, inoculating the cell cluster into an ultralow adsorption cell culture plate, and starting to culture, marking the time as the 0th hour of differentiation; and after culturing for 70-76 hours, replacing the basic differentiation culture solution containing the GSK-3beta inhibitor with a basic differentiation culture solution containing a Knockout TM serum substitute (KSR), and continuing to carry out differentiation culture. By adopting the method disclosed by the invention, the human induced pluripotent stem cells can be efficiently, quickly, simply, conveniently and stably subjected to directional induced differentiation into kidney organs containing podocytes, renal tubules and other structures, and the time for constructing a kidney in-vitro model can be shortened, so that the research on kidney diseases, kidney development and the like can be greatly assisted; and a powerful platform is provided for kidney toxicity assessment.

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 relates to the technical field of biology, and particularly discloses a construction method and application of an endometrial epithelium organ fibrosis model. According to the method, human endometrial epithelial cells are extracted, and an endometrial epithelial organ is established by adopting a Matrigel three-dimensional culture system. Afterwards, the organoid is induced to form endometrial phenotypes in different physiological cycles through hormone stimulation, and a ferroptosis inducer FIN56 is added in the culture process to induce fibrosis. The expression of GPX4, Fibronectin and p-Smad2 / 3 is detected through Western blot and immunofluorescence, and the successful construction of the fibrosis model is verified. The invention provides an in-vitro model capable of simulating endometrial fibrosis pathological characteristics of a patient with PCOS (polycystic ovarian syndrome), the in-vitro model can be used for mechanism research and drug screening of PCOS-related endometrial lesions, and a new research tool and an intervention strategy are provided for treatment of PCOS endometrial fibrosis.

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.

The invention provides a keloid disease in-vitro model as well as a construction method and application thereof, and relates to the technical field of cell biology. The construction method comprises the following steps: constructing a full-thickness skin model with an epidermal layer and a corium layer by utilizing a cell culture chamber or a cell culture micro-fluidic chip, and then performing induced culture by adopting TGF-beta to obtain the keloid disease in-vitro model. According to the construction method, by selecting the concentration and culture time of TGF-beta in the induction process, keloid-like change of the full-thickness skin model can be effectively promoted, and the pathological state of keloid is truly simulated; meanwhile, the human fibroblasts and the human keratinocytes are adopted to prepare the in-vitro model with the full-thickness skin structure, the in-vitro model has the human attribute, the individual difference of species difference bases is reduced, and the real effectiveness of the model is greatly improved; the defect that keloid which is a special human disease cannot be effectively simulated by an animal model is overcome.

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 present invention belongs to the field of biomedical technology and discloses an immortalized cell line of human renal chromophobe carcinoma, a culture method and an application thereof. The immortalized cell line of human renal chromophobe carcinoma of the present invention is named human renal chromophobe carcinoma cell line Loya-710 (Homo sapiens), and its Latin name is Chromophobe renal cell carcinoma:Loya‑ 710 , deposited with CCTCC NO: C2025123. The human chromophobe renal cell carcinoma cell line Loya-710, as an in vitro model, grows rapidly in tissue culture and retains the mitochondrial mutations, vesicle structure, and classic immunohistochemical markers characteristic of ChRCC. This invention provides a valuable tool for further studying the genetic, molecular, and biological characteristics of ChRCC and offers a powerful new model for mitochondrial disease.

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 present invention relates to an intestinal epithelium model faithfully reproducing the pathophysiological mechanisms observed in vivo in the context of an inflammatory state of the intestinal barrier. The model according to the present invention comprises two compartments separated by a semi-permeable membrane. The first compartment, corresponding to the apical pole of the intestinal epithelium, comprises a coculture of Caco-2 cells differentiated into enterocytes and HT29-MTX cells differentiated into goblet cells. The second compartment, corresponding to the basolateral pole of the intestinal epithelium, comprises a culture of THP-1 monocytic cells differentiated into macrophages.The model according to the present invention is characterized in that the cells contained in the first compartment produce interleukin (IL-6) at a concentration greater than 100 pg / mL, and interleukin 8 (IL-8) at a concentration greater than 150 pg / mL, and the cells contained in the second compartment produce tumor necrosis factor α (TNF-α) at ​​a concentration greater than 40 pg / mL and interleukin 1β (IL-1β) at a concentration greater than 90 pg / mL. The present invention also relates to the method for obtaining this model as well as to a method for selecting candidate compounds for the treatment of inflammatory bowel diseases by verifying their ability to stop inflammation upon increase in permeability or to verify the restoration of permeability and the mucus layer.

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.