79results about "Coculture" patented technology

The invention relates to a cell collection method capable of simultaneously collecting three cells in a co-culture model, and belongs to the technical field of biology. The invention provides a cell collection method capable of simultaneously collecting three cells in a co-culture model, and the cell collection method comprises the following steps: after a three-cell co-culture model is constructed, taking out a Transwell chamber, retaining cells in the lower chamber, and collecting the cells in the lower chamber; respectively digesting the cells on the two sides of the Transwell cell membrane by using a trypsin solution with the concentration of 0.5 g / 100mL so as to respectively collect the cells on the two sides of the Transwell cell membrane. According to the cell collection method disclosed by the invention, the three cells in the three-cell co-culture model are simultaneously collected in a manner of digesting the cells on the two sides of the Transwell membrane step by step by using pancreatin, so that not only is the cell and consumable cost saved, but also the experimental synchronism of the three cells is ensured, and convenience is provided for optimizing the experimental process.

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 relates to a retina nerve and blood vessel unit in-vitro three-culture model and a preparation method thereof, and belongs to the technical field of biology. The invention provides an in-vitro three-culture model of a retina nerve and blood vessel unit. The in-vitro three-culture model is obtained by carrying out in-vitro Transwell indirect co-culture on retina microvascular endothelial cells, retina ganglion cells and retina Muller cells. The in-vitro three-culture model of the retina nerve and blood vessel unit is based on a three-cell indirect co-culture method, retina microvascular endothelial cells, retina ganglion cells and retina Muller cells are simultaneously incorporated by utilizing Transwell, and the three cells are in indirect contact. The in-vitro three-culture model of the retina nerve and blood vessel unit can be used for more objectively simulating pathophysiological states of various cells of the retina nerve and blood vessel unit, so that common pathophysiological changes of mutual dependence and interaction of the three cells under in-vitro conditions can be observed.

The application discloses a kind of hepatic organoids containing mesenchyme induction method, belong to organoid culture technical field.The method includes the following steps: (1) human embryonic stem cell WIBR3 pretreatment;(2) utilize human embryonic stem cell WIBR3 and induce liver organoids and mesenchymal cells;(3) mesenchymal cells and liver organoid cells are co-cultured, i.e. obtain the hepatic organoids containing mesenchyme.There is beneficial effect: the application changes the culture mode and method of the hepatic organoids containing mesenchyme.The hepatic organoids containing mesenchyme induction method proposed in the application is simple, and mesenchymal cells and other liver cells have the same genetic background source, suitable for the safety evaluation of drug and other chemical substances, liver fibrosis, non-alcoholic hepatitis modeling and pharmacodynamic evaluation.

The invention discloses a preparation method and application of a conditioned culture solution for improving the development rate of bovine in-vitro embryos. The preparation method comprises the following steps: culturing immortalized bovine endometrial epithelial cells by using a DMEM / F12 cell culture solution containing 10% FBS, collecting the cell culture solution in the culture process, and filtering to obtain an immortalized bovine endometrial epithelial cell conditional culture solution, namely the conditional culture solution capable of improving the development rate of the bovine in-vitro embryos. The prepared conditioned culture solution can simulate a liquid environment of an in-vivo environment, is used for culturing in-vitro embryos such as parthenogenetic activated embryos and in-vitro fertilized embryos, improves the cleavage rate and blastocyst rate of the in-vitro embryos and improves the development rate of the in-vitro embryos of cattle.

Several embodiments disclosed herein relate to methods and compositions for enhanced expansion of NK cells in culture, through the repeated co-culturing of NK cells with feeder cells and the selective use of stimulatory interleukins. In several embodiments, the methods utilize one or more soluble interleukins as culture media supplements at one or more time points during expansion of the NK cell, or other immune cell, which results in a highly expanded and highly cytotoxic population of cells, for use in, for example allogeneic cellular immunotherapy.

The invention discloses a preparation method of a cochlear organ chip integrated with a blood labyrinth barrier, and belongs to the field of biomedical engineering. The preparation method of the simulated blood labyrinth barrier comprises the following steps: constructing an endothelial cell culture cavity and a pericyte culture cavity, and separating the endothelial cell culture cavity and the pericyte culture cavity through a simulated basement membrane; and respectively inoculating percutaneous cells and endothelial cells into a percutaneous cell culture cavity and an endothelial cell culture cavity, and culturing to obtain the simulated blood lost barrier. The chip can effectively simulate the structure and the function of the BLB, and the barrier integrity of the BLB is evaluated through TEER (trans-epithelial electrical resistance) measurement and an apparent permeability coefficient. The invention also relates to application of the chip in NIHL drug screening, in particular to drug evaluation for oxidative stress induced inner ear organ injury. Through the platform, the protection effect of the candidate drugs on the TBHP-induced oxidative stress injury of the inner ear organs can be evaluated, and a new tool and thought are provided for developing novel NIHL treatment drugs.

Provided is a method for comprehensive evaluation of the biological potency of mesenchymal stem cells, comprising: simulating in vitro a scenario where mesenchymal stem cells enter an inflammatory site in vivo, detecting expression levels of PD-L1, IL-6, HGF, and IDO1 in the mesenchymal stem cells, and detecting inhibition rates of proliferation of immune cells and secretion of inflammatory factor TNF-α, so as to evaluate the immunomodulatory function and tissue regeneration function of the mesenchymal stem cells.

The invention relates to the technical field of tumor model construction, and discloses a chick embryo allantoic membrane model (CAM)-based liver cancer tumor planting improvement method, which comprises the following steps: S1, preparing a tumor cell suspension; s2, carrying out nude mouse subcutaneous transplantation; s3, nude mouse subcutaneous tumor treatment; and S4, performing CAM planting. According to the chick embryo allantoic membrane model (CAM)-based liver cancer tumor planting improvement method, liver cancer cells are firstly cultured subcutaneously in a nude mouse to form tumor tissues, then the tumor tissues are cut into pieces and mixed with matrigel and a DMEM culture medium, the tumor tissues are planted in a chick embryo allantoic membrane, the growth state of the tumor tissues is optimized in the nude mouse subcutaneous environment, a bionic microenvironment is provided by a mixed solution, the tumor formation rate is increased, and the survival rate of the chick embryo allantoic membrane model (CAM) is increased. A silica gel ring is fixed at a bleeding point of the chick embryo allantoic membrane, tissue suspension is dropped, a preservative film with holes is covered for culture, the silica gel ring prevents the suspension from leaking out, the preservative film guarantees gas and nutrition exchange, the contact between tumor tissues and chick embryo allantoic membrane blood vessels is promoted, the tumor is larger, and observation and experimental operation are facilitated.

The present invention relates to systems, compositions and methods for multicellular bioprinting vascularized tissue constructs comprising a hierarchical vascular network. The methods employ a dual bio-ink strategy involving fibrous microgel matrix bio-inks with adjustable hardness, self-healing rheology, and microporosity to support capillary self-assembly, and sacrificial bio-inks that define a large pourable channel after removal. The system integrates top-down printing and bottom-up cell self-organization within a programmable bio-printing platform. The key aspect of the invention is to engineer the physical topography of the matrix to actively guide vascular morphogenesis beyond conventional permissive stents. The integrated platform enables the manufacture of complex bionic vascular networks with enhanced mass delivery, cellular viability, and in vivo integration potential. The invention represents an important progress in the aspect of extendable production of functional human tissues, and has application in regenerative medicine, disease modeling and drug discovery.

A method for culturing a human hepatic parenchymal cell, the method comprising culturing the human hepatic parenchymal cell in a culture medium comprising a culture supernatant of a human hepatic nonparenchymal cell, a Wnt signaling promoter and a cell-division-promoting growth factor.

The invention discloses an edible beefsteak scaffold based on vegetable protein and a construction method thereof, and belongs to the fields of tissue engineering, material science, food science and biology. The edible beefsteak scaffold is a 3D printing scaffold based on vegetable protein, combined with gellan gum and sodium alginate, and is a cell culture meat bionic beefsteak composed of a bovine fibroblast and bovine muscle satellite cell co-culture system dynamically constructed by using a rotary culture system. The selected materials are all edible protein and polysaccharide, have high biocompatibility and safety, and are wide in source; the mode of dynamically constructing the bionic beefsteak can effectively reduce the formation of local necrotic areas, and the bionic beefsteak has excellent physical characteristics, has texture structures and flavor characteristics similar to those of real beefsteak, is sterile in surface, and can significantly reduce food sanitation and safety problems. The scheme provided by the invention has remarkable advantages in the aspects of food safety, texture regulation and control, sensory simulation and the like, and provides important technical support for the field of cell culture meat.

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

Compositions and methods for stimulating the proliferation and cytotoxicity of γδ T cells are described. Therapeutic compositions and methods using the expanded and stimulated γδ T cells are described.

The invention provides a construction method and application of a sepsis model based on a lung and kidney organ chip. The method comprises the following steps: S1, carrying out ultraviolet sterilization on an organ chip device; s2, modifying a transwell plug-in unit; s3, inoculating human lung microvascular endothelial cells and human umbilical vein endothelial cells; S4, inoculating human alveolar epithelial cells and human renal tubular epithelial cells; s5, transwell inserts are arranged in two holes in the middle of the organ chip device respectively, a mixed culture medium is added into liquid adding holes in the two sides of the organ chip device, and the lung chip and the kidney chip are communicated through a lower channel of the organ chip device; s6, dynamically culturing the lung and kidney organ chip; and S7, sucking up the culture medium in the constructed lung and kidney organ chip, adding the LPS-containing mixed culture medium into a lower channel, and respectively adding the LPS-containing mixed culture medium into the human alveolar epithelial cell side and the human renal tubular epithelial cell side of the lung and kidney organ chip and the transwell plug-in hole for culturing for 2 days. According to the invention, the lung and kidney organ chip is applied to the sepsis model for the first time, and the constructed sepsis model is more suitable for basic research and drug development of sepsis.

Provided is an embryo model, which is obtained by means of self-assembly of induced pluripotent stem cells and induced hypoblast stem cells, wherein both the induced pluripotent stem cells and the induced hypoblast stem cells are obtained by means of somatic cell reprogramming. The self-assembly of the stem cells of two different lineages generated by means of somatic cell reprogramming forms an embryo model simulating human peri-gastrulation. The embryo model better reproduces the interaction and communication mechanism among cells of multiple lineages during human embryonic development by means of combining cells of different lineages. With the embryo model, by means of a gene editing tool, lineage tracing and gene regulatory network research can be performed on cells of different lineages, thereby enabling an in-depth understanding of the molecular regulatory mechanisms of early human embryonic development.

The invention discloses an ovarian cancer organoid-macrophage co-culture system as well as a preparation method and application thereof. The preparation method comprises the following steps: providing ovarian cancer cells; the method comprises the following steps: resuspending ovarian cancer cells in matrigel, then inoculating the matrigel on which the ovarian cancer cells are resuspended into a culture container, after the matrigel is completely solidified, adding the matrigel into an ovarian cancer organoid complete culture medium, culturing to obtain an ovarian cancer organoid wrapped in the matrigel, and separating to obtain the ovarian cancer organoid; providing macrophage M2, mixing the macrophage M2 and the ovarian cancer organoid, and culturing by using a co-culture medium. In combination with a specific embodiment, according to the preparation method of the ovarian cancer organoid-macrophage co-culture system, macrophages M2 and ovarian cancer organoid are mixed and then cultured with a co-culture medium, the obtained ovarian cancer organoid-macrophage co-culture system contains the macrophages M2, and the functions of the macrophages can be researched.

The invention discloses an ovarian cancer organoid-T cell co-culture system as well as a preparation method and application thereof. The preparation method comprises the following steps: providing ovarian cancer cells; the method comprises the following steps: resuspending ovarian cancer cells in matrigel, then inoculating the matrigel on which the ovarian cancer cells are resuspended into a culture container, after the matrigel is completely solidified, adding the matrigel into an ovarian cancer organoid complete culture medium, culturing to obtain an ovarian cancer organoid wrapped in the matrigel, and separating to obtain the ovarian cancer organoid; t cells are provided, and the T cells and the ovarian cancer organoid are mixed and then cultured with a co-culture medium. In combination with a specific embodiment, according to the preparation method of the ovarian cancer organoid-T cell co-culture system, T cells and ovarian cancer organoid are mixed and then cultured by a co-culture medium, the obtained ovarian cancer organoid-T cell co-culture system contains the T cells, and T cell immune response can be truly reflected.

The present invention relates to a pathological pulmonary fibrosis model, a construction method therefor, and a use thereof. The pathological pulmonary fibrosis model provided by the present invention has good stability and controllability and high maturity, can better simulate the function and structural characteristics of the alveolar-capillary barrier, and has the pathological characteristics of pulmonary fibrosis lesions and the immune microenvironment. The model has the advantage of high simulation degree, and has good application prospects in the aspects of simulation and research of pulmonary fibrosis diseases, as well as drug detection, screening, assessment, and research and development, and the like.

The invention provides a smoke disease model and a construction method and application thereof, and belongs to the technical field of smoke disease model construction. The method for constructing the smog disease model comprises the following steps: extracting CD34 + hematopoietic progenitor cells in peripheral blood of a smog disease patient, reprogramming to generate hiPSC, further differentiating into brain organs and vascular organs, fusing the brain organs and the vascular organs, and culturing and constructing the vascularized brain organ smog disease model. The smoke disease model constructed by adopting the construction method disclosed by the invention can simulate and reproduce phenotypes such as tiny branch increase, macrovascularization, pericytopenia, smooth muscle fibrosis deposition, abnormal angiogenesis increase and the like in clinical core pathological phenotypes of smoke disease patients, and is superior to existing animal and cell models to a certain extent.

Provided are an induced NK cell, and a preparation method therefor and the use thereof. The method comprises mixing a CD34+ cell with a stromal cell, co-culturing the mixture to obtain an NK precursor cell, and subjecting the NK precursor cell to induction and amplification culture to obtain a mature NK cell, wherein the stromal cell comprises any one or a combination of at least two of an AFT024 cell, an MS5 cell, an OP9 cell, an HS-5 cell, an MSC cell, an MUTZ-3 cell, a stromal cell derived from tissues such as bone marrow or the liver, or a primary cell. Provided is the development of a brand-new method for inducing an NK cell on the basis of a CD34+ cell, which enables the induction of the NK cell with a high efficiency. The efficiency of outputting NK cells from a single CD34+ cell is improved, and the property of the induced NK cell is very close to that of a natural mature NK cell. They have uniform anti-tumor effects, can effectively kill tumor cells, and have broad application prospects.

The invention belongs to the technical field of biomedical engineering and microfluidics, and particularly relates to a vascularized organ chip capable of collecting multi-cell co-culture or organoid metabolites in situ for multiple times. The invention provides a vascularized organ chip. The vascularized organ chip sequentially comprises a runner layer, a cell culture layer, a porous membrane layer and a metabolite collection layer, the runner layer is provided with a culture solution runner, and the culture solution runner is communicated with the outside through a culture solution hole; the cell culture layer is provided with a plurality of stem cell culture chambers; each cell culture chamber is communicated with the outside through the hydrogel channel and the cell ball injection hole; the area of the porous membrane layer at least covers the bottoms of all the cell culture chambers; the metabolite collecting layer is provided with a plurality of collecting chambers, and at least one collecting chamber is arranged below each cell culture chamber; the vascularization organ chip is further provided with a plurality of liquid taking channels, and each collecting cavity is communicated with the outside through the corresponding liquid taking channel. The vascularized organ chip provided by the invention can realize in-situ collection and dynamic analysis of metabolites.

Embodiments of the instant disclosure relate to novel compositions and methods for generating thymic cells. In some embodiments, thymic cells can be differentiated from pluripotent stem cells (PSC), anterior primitive streak (APS) cells, definitive endoderm (DE) cells, anterior foregut endoderm (AFE) cells, pharyngeal endoderm (PE) cells, ventral pharyngeal endoderm (VPE) cells, and third pharyngeal pouch endoderm (TPPE) cells using the compositions and methods disclosed herein. In certain embodiments, thymic cells generated by composition, systems and methods disclosed herein can be used to treat a health condition.

A technology regarding the production, formulation and use of conditioned media and the factors included therein is disclosed. The conditioned media may be inoculated with animal cells, plant cells and any combination thereof. The inoculations may occur simultaneous or at different times. Cells retrieved from different areas of the animal and / or the plant may also be cultured together to form conditioned media and associated growth factors.

This invention discloses a culture medium and method for co-culturing neural stem cells and microglia, belonging to the field of cell co-culture technology. It includes a basal culture medium, a dual-mode liposome, and the following components: B-27 serum-free additive (vitamin A-free), GlutaMAX additive, N-acetylcysteine, and ascorbic acid-2-phosphate; wherein the dual-mode liposome includes growth factor liposomes and pH-responsive liposomes. This invention addresses the core pain point of co-culturing neural stem cells and microglia, overcoming the concentration fluctuation defects of traditional pulsed addition of free factors. It develops a dual-mode liposome delivery system, which improves the stability of cytokine action, solves the concentration fluctuation problem caused by traditional pulsed addition of free factors, achieves stable delivery, and significantly improves the survival rate of co-cultured cells.

Provided in some aspects are methods for generating primordial follicles from human and other mammalian germ cells, with high efficiency, high cellular quality and high reproducibility. In some aspects, provided are methods and culture system for generating ovarian organoids, such as cell aggregates composed of both somatic and germ cells from primary and / or stem cell-derived sources, which are competent to progress primordial germ cells to oocytes that enter meiosis and are competent for follicle assembly. In some aspects, provided embodiments include a step to culture ovarian organoids in a fully defined serum-free media and in conditions with low defined serum protein components overall.

This invention belongs to the field of organ-on-a-chip technology, proposing a method for constructing an in vitro brain lymphatic system chip and a brain lymphatic system chip, filling the gap in the construction of in vitro models of brain lymphatic systems. First, the microfluidic channels in the chip are rinsed, sterilized, washed, and cultured with cells. Then, vascular channel cells and brain parenchyma channel cells are seeded in the treated microfluidic channels. Finally, in the vascular channels, perfusion is performed at a preset flow rate and at preset intervals to provide shear force stimulation mimicking pulsed blood flow. Continuous perfusion achieves the basic formation of in vitro brain blood vessels. The intermittent fluid flow mimicking arterial pulsations is achieved through this preset flow rate and preset interval perfusion.

This invention relates to a printing apparatus using multiple inks and a printing method using the same apparatus. More specifically, the invention relates to a printing apparatus using multiple inks, the printing apparatus comprising an ink extrusion member, the ink extrusion member comprising: a storage member divided into a plurality of receiving members for respectively receiving multiple inks; and an extrusion member having a single channel through which the multiple inks received in the storage members pass. The invention also relates to a three-dimensional printing method for producing a printed output with a cross-sectional pattern, the method comprising the steps of: using the printing apparatus to supply different inks to the divided spaces and applying the same pressure conditions to the received inks, so as to extrude the inks received in two or more divided spaces through a single extrusion port, thereby producing an ink extrusion product and printing.

The application provides a method for constructing a transplanted tumor model based on macrophage co-culture, and belongs to the technical field of biotechnology.The method aims to solve the problems of low success rate, slow growth rate and drug resistance of traditional transplanted tumor model construction, and significantly improves the construction efficiency and tumor growth rate of the transplanted tumor model by introducing a macrophage co-culture step, which is helpful for tumor drug screening and efficacy prediction.

The invention discloses a method for obtaining enhanced IMD-0354 and application of the enhanced IMD-0354 in neuroprotection, and relates to the technical field of biological medicine. The method comprises the following steps: 1) regulating and controlling the expression or activity of IRF5 in myeloid-derived proinflammatory immune cells, so that the expression quantity of the IRF5 is reduced or the activity of the IRF5 is inhibited; and (2) co-culturing the IRF5 regulation type myeloid-derived proinflammatory immune cells obtained in the step (1) and nerve cells in a neurodegenerative disease cell model related to TDP-43 pathology in a culture system containing IMD-0354 so as to enhance the protection effect of the IMD-0354 on the nerve cells. The application disclosed by the invention has the beneficial effects that the neuroprotection of the IMD-0354 on ALS cells is enhanced, the interference of proinflammatory macrophages on the IMD-0354 is antagonized, and a thought is provided for ALS treatment and drug research and development.