107 results about "Co-Culture" patented technology

This invention relates to the field of plant genetic engineering technology, and more particularly to... SsNHX1 and SsSOS2 Dual-gene plant expression vector and its application in improving salt tolerance in alfalfa. The plant expression vector uses pCAMBIA1300 as its backbone and inserts genes derived from Suaeda salsa. SsNHX1 Genes and SsSOS2 It is constructed from genes. SsNHX1 Genes and SsSOS2 The CDS coding sequence of the gene is shown in SEQ ID NO:1 and SEQ ID NO:2 of the sequence listing. Using Agrobacterium tumefaciens EHA105-mediated transformation, the vector was introduced into the leaves of sterile alfalfa seedlings. Positive transgenic plants were obtained through co-culture, callus induction differentiation, and PCR identification. The advantage lies in the synergistic effect of the two genes, which effectively enhances the salt tolerance of alfalfa, enabling it to grow normally under 200–300 mM NaCl stress, providing a new solution for forage cultivation in saline soils.

This invention relates to the field of plant genetic engineering technology, and more particularly to... SsNHX1 and SsSOS1 A dual-gene plant expression vector and its application in improving salt tolerance in alfalfa. The plant expression vector uses pCAMBIA3301 as its backbone and inserts genes derived from Suaeda salsa. SsNHX1 Genes and SsSOS1 It is constructed from genes. SsNHX1 Genes and SsSOS1 The nucleotide sequences of the gene are shown in SEQ ID NO:1 and SEQ ID NO:2. Using Agrobacterium tumefaciens EHA105-mediated transformation, the vector was introduced into the leaves of sterile alfalfa seedlings. Positive transgenic plants were obtained through co-culture, callus induction differentiation, and PCR identification. The advantages are: the synergistic effect of the two genes effectively enhances the salt tolerance of alfalfa, enabling it to grow normally under 200–300 mM NaCl stress, providing a new solution for forage cultivation in saline soils.

This invention relates to the fields of biomedical technology and biological model construction technology, and discloses an ANCA-associated vasculitis glomerular endothelial cell injury model, its construction method and application, and the application of BACH1 inhibitors. The method includes: (1) activating neutrophils sequentially with tumor necrosis factor α and immunoglobulin G to obtain a culture medium containing activated neutrophils; wherein the immunoglobulin G is immunoglobulin G isolated and purified from the plasma of patients with ANCA-associated vasculitis; (2) co-culturing the culture medium containing activated neutrophils with human glomerular endothelial cells to obtain the cell injury model. The cell injury model provided by this invention can verify the role of neutrophil adhesion characteristics in endothelial cell injury, and at the same time, the cell injury model can better simulate the immune microenvironment of ANCA-associated vasculitis glomerular endothelial cells in vivo.

The invention relates to a method to independently identify by fluorescence microscopy different cell types forming multicellular structures in co-culture thanks to a fluorescently-labelled extracellular matrix, and the individual identification of each multicellular structure of the co-culture with a spatial positioning of those structures in microcavities, so as to individually and independently measure multiple biological activities in each multicellular structure of each type of tissue of the co-culture.

Provided are methods for expanding tumor-infiltrating lymphocytes (TILs), which include co-culturing an initial cell population containing TILs with first feeder cells to obtain a first expanded cell population; and then co-culturing the first expanded cell population with second feeder cells to obtain an expanded TIL population. The methods can quickly produce a large number of TILs from a small tumor sample.

This invention discloses a T-cell training method based on tumor organ-on-a-chip. The method includes: preparing an organ-on-a-chip with a micropillar array and a biomimetic fishbone structure; introducing primary tumor cells from a patient into the chip, causing them to spontaneously form uniform three-dimensional tumor spheroids; dynamically pumping pre-activated peripheral blood lymphocytes from the same patient into the chip and co-culturing them with the tumor spheroids; collecting the co-cultured lymphocytes and repeating the co-culture process multiple times to obtain a lymphocyte population with enhanced anti-tumor activity. This invention utilizes organ-on-a-chip technology to highly simulate the dynamic tumor immune microenvironment in vivo. Through multiple cycles of "training," it effectively activates and expands tumor-specific T cells, avoiding immune exhaustion, and providing a highly efficient and controllable new platform for personalized immunotherapy of solid tumors.

This invention relates to a genetic transformation method for Ningxia wolfberry using germinating seeds as explants, belonging to the field of plant genetic transformation technology. The invention mainly includes the following steps: (1) After disinfecting Ningxia wolfberry seeds, inoculate them onto a germination medium, and obtain germinating seeds through vernalization and dark culture treatment; (2) Activate and culture Agrobacterium rhizogenes and Agrobacterium tumefaciens strains carrying target gene expression frames to obtain a bacterial solution for infection; (3) Take the germinating seeds obtained in step (1) and mix them with the bacterial solution obtained in step (2) for infection, then transfer the infected germinating seeds to a co-culture medium or substrate for co-culture; (4) Cultivate the material after co-culture in step (3) to induce root growth, and detect the expression level of the target gene or reporter gene on the roots or plants to obtain transgenic positive plants. This invention has the advantages of simple operation, significantly shortened transformation cycle, and high transformation efficiency.

The preparation method of the biomimetic stratum corneum and the biomimetic skin with tissue-like differentiation structure belongs to the technical field of transdermal drug delivery. Polymer microspheres are dispersed in a lipid mixture to obtain lipid-coated microspheres, and then a polymer-lipid composite film with a brick-mortar-like structure is formed by hot pressing, thereby obtaining a biomimetic stratum corneum highly consistent with the structure of human stratum corneum. The biomimetic stratum corneum not only has anisotropic ultrastructure in the natural stratum corneum, but also shows a permeation barrier function highly consistent with the ex vivo human skin. The biomimetic stratum corneum is co-cultured with an artificial tissue culture skin to prepare a biomimetic skin, so as to introduce skin metabolic function and / or serve as a hair follicle stem cell carrier, and construct a biomimetic skin containing a hair follicle structure. The present application is suitable for in vitro transdermal penetration test and evaluation of drugs and cosmetics, and provides reliable and efficient alternative materials and methods to solve the problems of poor predictability and large individual differences of existing animal skin models.

The application provides a trophoblast organ-on-chip system for simulating a maternal-fetal interface of a systemic lupus erythematosus model based on an organ chip and a construction method thereof, the system taking an organ chip as a carrier and comprising three core parts of a trophoblast organ, an endometrial chip and an SLE disease model. The application completes separation culture and characterization of the trophoblast organ, multi-cell co-culture construction and characterization of the endometrial chip, and then constructs a pathological model, and the three parts cooperatively form a three-dimensional maternal-fetal interface. The system can clearly observe the dynamic process of the trophoblast organ invading the endometrial chip, can detect the continuous secretion of hCG and SLE-related inflammatory factors, and breaks through the defects of the prior art model, such as low authenticity, inability to dynamically observe, single detection dimension and lack of standardization, and has both physiological and pathological adhesion and experimental repeatability, can multi-dimensionally characterize the cell phenotype and function of the maternal-fetal interface, and provides an in-vitro research platform for the pathological mechanism research of the SLE pregnancy-related abnormal maternal-fetal interface.

This invention belongs to the field of biomedical engineering technology, specifically the field of cell function detection technology, and relates to a method for evaluating the ability of mesenchymal stem cells (MSCs) to promote Treg cell proliferation. This method utilizes porous microcarriers to achieve three-dimensional cell distribution, employing low-speed rotation throughout the culture process to simulate the in vivo suspension microenvironment. A culture medium matching human physiological parameters is configured, with oxygen concentration adjusted to a hypoxic range of 1%-3% to simulate the microenvironment of inflammatory sites. Mononuclear cells with a purity of over 95% are obtained through magnetic bead sorting and co-cultured with MSCs for approximately 3 days, significantly shortening the culture period and reducing data deviation, thus achieving specific screening of MSCs with high immunomodulatory functions. By constructing a co-culture microenvironment that conforms to human physiological characteristics, this invention effectively improves detection sensitivity and repeatability, avoids interference from individual differences in peripheral blood mononuclear cells from different sources, and provides technical support for the quality control and clinical application of MSCs.

This invention relates to the field of cell biology, specifically to a co-culture cell model and method for primary glial cells and neurons. The method for co-culturing primary glial cells and neurons provided by this invention includes the following steps: a mixture of primary glial cells and neurons is seeded into a first co-culture medium for pre-culture; the medium is discarded and the cells are washed; a second co-culture medium is added for a first-stage culture; after the first-stage culture, a second-stage culture is performed; the medium used in the second-stage culture is a third co-culture medium supplemented with macrophage colony-stimulating factor; after the second-stage culture, a third-stage culture is performed; the medium used in the third-stage culture is the second co-culture medium; in the obtained co-culture cell model, microglia are differentiated and mature, and are in a quiescent state, thus obtaining a primary glial cell and neuron co-culture model.

The application provides a corn genetic transformation method for obtaining a homozygous event in a tissue culture current generation, comprising the following steps: (1) using agrobacterium containing a target gene to infect a corn inbred line ND101 haploid embryo, to obtain an infected haploid embryo; (2) co-culturing the infected haploid embryo, to obtain a corn inbred line ND101 haploid embryo after co-culture; (3) sequentially performing resistance callus induction and doubling treatment on the corn inbred line ND101 haploid embryo after co-culture, to obtain a resistance callus; (4) sequentially performing pre-differentiation culture, differentiation culture and rooting culture on the resistance callus, to obtain a transgenic positive T0 generation homozygous diploid plant. The method combines genetic transformation and haploid technology through a specific culture medium, realizes one-step obtaining of a homozygous event, and shortens the traditional 2-3 generations to be completed within one generation.

This invention belongs to the field of biomedicine and tumor immunotherapy technology, specifically relating to a monoclonal antibody targeting the immune checkpoint molecule CD276 (B7-H3) and its applications. The applicant conducted immunization and screening work around the human CD276 antigen, selecting two representative monoclonal antibody clones, 2B2-1 and 5G4-2, from multiple candidate antibodies. The nucleotide and amino acid sequences of their heavy and light chain variable regions were determined, and their performance in flow cytometry, T-cell killing assays, immunohistochemistry, ELISA, and Biacore affinity assays was systematically evaluated. Experimental results show that the above antibodies can specifically recognize and bind to CD276, efficiently distinguish between CD276-high and low-expressing cells, and enhance the killing activity of T cells against CD276-high expressing tumor cells in an in vitro co-culture system. This provides an excellent antibody backbone for the subsequent construction of diagnostic kits, ADC drugs, or bispecific antibodies.

This invention provides a method for transient transformation of oil palm zygote embryos based on Agrobacterium-mediated transformation, which employs the Agrobacterium-mediated approach and is based on the optimal Agrobacterium concentration (OD) for stable transformation of oil palm. 600 The optimal Agrobacterium infection time for zygotic embryos was determined by measuring the concentration of acetylsuccinone and co-culture time. The transient GUS expression staining rate was compared between zygotic embryos of different oil palm varieties and at different growth stages. The method described in this invention optimizes infection conditions based on stable transformation parameters, clarifies the optimal explant materials and treatment process, and effectively improves transient transformation efficiency and experimental repeatability. This study establishes for the first time an Agrobacterium-mediated transient transformation system for oil palm zygotic embryos and obtains zygotic embryos most suitable for transient transformation, achieving efficient transient expression of exogenous genes in oil palm zygotic embryos. This provides a rapid and low-cost technical means for studying oil palm gene function.

The application discloses a traditional Chinese medicine exosome compound with anti-HPV and application thereof, and relates to the technical field of biotechnology.The compound comprises functional components secreted by a human cell co-culture system and a traditional Chinese medicine source extract, and the traditional Chinese medicine source extract is derived from a combined traditional Chinese medicine extract of Astragalus membranaceus, Isatis indigotica Fort., Honeysuckle and Sophora flavescens Ait., and relates to the technical field of biotechnology.The traditional Chinese medicine exosome carries natural active ingredients, and a multi-target anti-virus network is formed by beta-defensin-2 and interferon-gamma generated by co-culture with human cells.Compared with single traditional Chinese medicine extract or chemically synthesized nanoparticles, the HPV inhibition rate is increased by more than 170%, and the direct cytotoxicity of traditional Chinese medicine is reduced through an exosome delivery system, and the IC 50 >200ug / mL, which is significantly lower than that of a water decoction, and mucous membrane irritation experiments show no damage.

The application relates to a tumor organoid-immune cell co-culture method, a co-culture system and application thereof. The method comprises the following steps: S1. adding pre-cooled mixed matrix material into a culture container, and forming a matrix layer after solidification; wherein the mixed matrix material comprises Matrigel matrix glue and gelatin hydrogel in a volume ratio of 1: (0.1-10); S2. inoculating tumor organoid suspension obtained through pretreatment onto the surface of the matrix layer, so that the tumor organoid is attached to the matrix layer, and dropping immune cell suspension obtained through pretreatment onto the surface of the tumor organoid attached to the matrix layer; and S3. adding co-culture medium into the culture container, so that the tumor organoid and the immune cell are immersed in the co-culture medium, and dynamic co-culture is carried out. The method solves the problems of poor co-culture compatibility, poor stability and low repeatability in the prior art, and realizes long-term, efficient and stable co-culture of the tumor organoid and the immune cell.

The embodiment of the application discloses a kind of vein bionics microfluidic chip and its preparation and application method of simulating multi-organ interconnection.The chip includes a chip main body, and its microfluid channel system integrates at least three organ simulation units: a lung simulation chamber for culturing lung cells to simulate upstream organs that can produce systemic effects;An upstream colon cancer cell culture chamber for three-dimensional culture of colon cancer cells to simulate tumor primary lesions;And a downstream liver cell culture chamber for three-dimensional co-culture of liver cells and vascular endothelial cells to simulate the target organ of liver, and the three simulation chambers are communicated with each other by a vein bionics microfluid network, which simulates the complex blood circulation system of human body (including pulmonary circulation and systemic circulation), can realize the signal molecule transmission between organs, and simulate the invasion and metastasis process of colon cancer cells to the target organ of liver via the circulatory system after being affected by upstream lung signals.

The application discloses a di(2-ethylhexyl) phthalate degrading bacterium, a bacterial composition and application thereof. The degrading bacterium is named Sphingomonas parapaucimobilis GHQ28 and Stenotrophomonas maltophilia GHQ31. Research finds that GHQ28 and GHQ31 have chemotaxis to DEHP and downstream metabolites thereof, and there is a synergistic interaction between the two bacteria; the growth and DEHP degradation speed of the two strains under co-culture conditions are faster than that under single culture conditions. The bacterial composition composed of the two strains has a highest degradation rate of 99.2±0.59% to 400mg / L DEHP within 72h, compared with single culture, the total biomass is increased by 1.4-50.3 times, and the DEHP degradation rate is increased by 1.07-58.82 times. The application provides a new technical means for remediation of DEHP contaminated soil or water.

A device includes a first unit for skeletal muscle tissue formation; a second unit for motor neuron culture; a third unit for causing the first and second units to communicate with each other; and a pillar serving as a scaffold for skeletal muscle tissue formation. The first unit includes a first base material and a first culture tank formed in the first base material. The second unit includes a second base material and a second culture tank formed in the second base material. The third unit includes a third base material and an axon channel formed in the third base material, through which a bundle of axons passes. One end of the third unit is connectable to the second unit and cause the axon channel and the second culture tank to communicate with each other. A first opening part is formed to the other end of the third unit.

This invention discloses a high-throughput screening method based on cytokinesis for bidirectional recognition of TCR and pMHC. The method involves displaying both pMHC and TCR on the cell membrane, with the TCR simultaneously fused with the MS2 phage capsid protein for capturing barcode and MS2 RNA. The two cell types are co-cultured, and cytokinesis-mediated membrane transfer occurs between cells exhibiting bidirectional TCR and pMHC recognition. Cells displaying pMHC are isolated and collected for single-cell transcriptome sequencing analysis. The sequencing results are grouped according to different pMHCs, and the corresponding TCR is determined using the barcode sequence, thus identifying the TCR that specifically recognizes pMHC bidirectionally. This high-throughput screening method based on cytokinesis for bidirectional specific recognition of TCR and pMHC overcomes, to some extent, the limitations of existing technologies in terms of screening throughput, physiological relevance, and bidirectional matching ability.

This invention discloses a co-culture immune isolation device after cell transplantation, belonging to the technical field of cell culture devices. The device includes a culture plate with several first cavities inside and placement slots outside the first cavities. A cover plate is provided above the culture plate, and several second cavities are provided on the cover plate. A sealing ring is provided on the outside of the second cavities, and a liquid inlet mechanism is provided inside the cover plate, connected to the second cavities. This invention uses the above-mentioned co-culture immune isolation device after cell transplantation to achieve co-culture of multiple cells by setting multiple independent first cavities, preventing cell migration or direct contact and avoiding immune rejection. The protective cover and liquid inlet mechanism achieve closed-loop automatic liquid inlet, continuously diffusing signal molecules into the first cavities to maintain co-culture homeostasis and achieve co-culture of multiple cells.

A co-culture model of brain glioma organoids and mouse brain slices, characterized in that the co-culture model is prepared by the following method: (1) taking mouse brain slices, culturing to obtain cultured mouse brain slices; (2) injecting brain glioma organoids into the cultured mouse brain slices obtained in step (1) to obtain brain slices injected with brain glioma organoids; (3) culturing the brain slices injected with brain glioma organoids obtained in step (2) to obtain the co-culture model. The co-culture model of the application can be used to evaluate the invasion phenotype of glioma organoids in vitro and to test and screen brain glioma drugs.

The invention relates to a recombinant microbial strain for de novo synthesis of gardenin B and application of the recombinant microbial strain in production of polymethoxylated flavone compounds, and belongs to a cross technology of synthetic biology and organic synthetic chemistry. The invention designs and constructs a heterozygous gardenin B biosynthesis pathway, and obtains a microbial strain for synthesizing gardenin B from the beginning. By screening F8H and redox partner types and regulating and controlling subcellular localization, an F8H-redox partner combination with optimal space and type is determined, and an engineering strain of which the gardenin B yield is greatly improved is obtained. The efficient synthesis method of the polymethoxylated flavonoids is established by selecting recombinant microorganisms for synthesizing gardenin B from de novo to perform co-culture fermentation or chemical conversion of fermentation products.

Featured are methods, compositions, and apparatuses for the in vitro maturation of oocytes. In particular, the disclosure features methods of inducing oocyte maturation in vitro, by co-culturing a female subject's oocytes with an ex vivo composition containing a plurality of ovarian support cells (e.g., granulosa cells). Additional methods for administering follicular triggering agents and retrieving oocytes from the female subject are provided. Such methods, compositions, and apparatuses are particularly useful for assisted reproduction technology (ART) procedures.

This invention belongs to the field of biology and discloses a method for co-culturing enteric glial cells (EGCs) and intestinal organoids. Primary cultured and purified enteric glial cells are digested and seeded onto the membrane of a Transwell chamber. Intestinal organoids are placed in individual Matrigel drops in a co-culture plate. The Transwell chambers are placed in the co-culture plate, and growth medium is added to ensure free exchange of the growth medium through the Transwell membrane. The EGC-organoid co-culture system obtained based on this method provides a model for studying diarrhea and intestinal health in large mammals. It also lays the research foundation for developing innovative intervention strategies targeting the enteric nervous system and provides a research model for further exploring the regulatory roles of nutrients or related genes in intestinal health.

The invention discloses a co-culture method of cell culture meat and application of the co-culture method in improving meat quality and flavor. According to the invention, through two-stage differentiation, muscle precursor cells and fat precursor cells are respectively subjected to proliferation culture, then are separately subjected to primary differentiation, and are mixed together for co-culture, and meanwhile, second-stage continuous differentiation is carried out; the problem that differentiation of muscle cells and fat cells cannot be effectively and simultaneously differentiated under the co-culture condition due to different requirements of differentiation of muscle cells and fat cells on the serum environment is solved. In addition, the invention also provides an objective standard for evaluating the flavor quality of cell culture meat generated by co-culture. The cell culture meat with different flavors can be obtained by adjusting the proportion of the co-culture cells through the co-culture method, and the flavor quality of the cell culture meat from other sources can be evaluated and detected through the amino acid detection means.

This invention relates to the field of agricultural biological control technology, specifically to a synthetic microbial co-culture fermentation broth, its inoculum, and its application. The synthetic microbial co-culture fermentation broth is prepared by sequential co-inoculation fermentation of *Trichoderma harzianum* TW21990, *Burkholderia viminalis* P418, and *Bacillus atrophicus* BsR05; wherein *Trichoderma harzianum* TW21990 has the CGMCC No. 12864, *Burkholderia viminalis* P418 has the CGMCC No. 1212, and *Bacillus atrophicus* BsR05 has the CGMCC No. 11665. Compared to single biocontrol strains, the antibacterial and growth-promoting secondary metabolites produced by the above-mentioned synthetic microbial co-culture fermentation are broad-spectrum, making it less likely to induce drug resistance in the control of gray mold in leeks. The control effect is stable, there is no antagonism within the synthetic microbial community, and the efficacy of the strains is synergistically enhanced, with a significant growth-promoting effect on leeks.

This invention discloses a microfluidic chip and its fabrication method for constructing modular cell cultures, relating to the field of cell culture technology. The chip includes an upper plate and a lower plate. The upper plate includes a first flow channel and a second flow channel, while the lower plate includes multiple spaced-apart culture chambers, each with a different cross-sectional shape. This allows each chamber to be independently formed into cultures of different shapes. The cultures can be stacked and combined after removal to construct a multi-cell co-culture model with functional partitions, significantly improving structural complexity and biomimicry. The upper plate slides to switch the connection between the first and second flow channels and the culture chambers, enabling inoculation and culture modes. During inoculation, cultured cells are injected into the culture chamber through the first flow channel; during culture, the flow channel is switched to perfuse the culture chamber with culture medium, avoiding damage to the culture during residual gel removal. This simple operation improves the success rate and enhances the reliability of experimental results.

The application provides a multifunctional microalgae-based hydrogel wound dressing and a preparation method and application thereof, and belongs to the field of hydrogel dressings. Microalgae are embedded in a hydrogel network formed by modified gelatin and cross-linked sodium alginate, and a multifunctional alginate hydrogel is constructed for chronic wound repair of diabetes. The selected microalgae can realize in-situ synthesis and uniform distribution of small-sized gold nanoparticles on the surface thereof during co-culture with high-concentration gold ions, stimulate high-efficiency secretion of extracellular polysaccharides by the microalgae, and maintain high cell activity. The prepared dressing is helpful for relieving a local hypoxic microenvironment of a wound and promoting vascular regeneration, can effectively remove excess active oxygen at a wound site, and can induce macrophages to polarize to M2 type; in addition, the gold nanoparticles in the hydrogel network have good antibacterial performance.