83 results about "Cell soma" patented technology

The invention relates to the technical field of biology, in particular to a Treg cell amplification culture medium and a preparation method of Treg cells. According to the method for in-vitro amplification of the regulatory T cells (Treg) from peripheral blood, the amplification efficiency and purity of the Treg cells are improved, the culture cost is reduced, the culture period is shortened, and the problem of trophoblast cell residues in subsequent clinical application is solved.

The invention belongs to the technical field of biological medicines, and particularly relates to an antigen peptide library for inducing tumor specific immune response, the antigen peptide library comprises eight antigen peptides, and the amino acid sequences of the eight antigen peptides are respectively shown as SEQ ID NO.1-SEQ ID NO.8. The invention further discloses a preparation method of the antigen peptide library. The antigen peptide has high immunogenicity and can effectively stimulate and induce generation of specific cytotoxic T lymphocytes when being loaded on antigen presenting cells, in-vitro killing experiments show that CTL cells induced and activated by loading new antigen DC have a high tumor killing effect, and the cell killing activity of the CTL cells is 90% or above. When model mice are treated by adoptive T cell back-transfusion of CTL cells induced by the new antigen peptide library, the tumor volume can be effectively reduced, the survival rate of the tumor can be remarkably increased, and good treatment potential is achieved.

The invention relates to a target cell in-vitro sorting method for cell therapy, and belongs to the technical field of cell therapy. According to the method, the cleaning solution containing EGTA, EDTA, alpha-casein and prostaglandin is used, so that the sorting method with high living cell recovery efficiency is obtained; and cell therapy can be better realized.

The invention belongs to the technical field of culture media, and particularly relates to a culture medium for promoting in-vitro amplification of NK cells, the culture medium for in-vitro amplification of the NK cells comprises a basic culture solution, a CD16 antibody, IL2, IL12 and IL21, the basic culture solution is DMEM, and the culture medium for in-vitro amplification of the NK cells is obtained by adding autologous plasma, the CD16 antibody, the IL2, the IL12 and the IL21 on the basis of the basic culture solution. Wherein the addition amount of the autologous plasma is 10%-15%, and the final concentrations of the CD16 antibody, IL2, IL12 and IL21 are 1 [mu] g / mL to 20 [mu] g / mL, 1000 IU / mL to 2000 IU / mL, 20 ng / mL to 40 ng / mL and 5 ng / mL to 15 ng / mL. According to the method, the culture medium is optimized, the single NK cell is resuspended by using the complete culture medium, the single NK cell is added into the CD16 antibody coated cell culture plate and cultured for 21 days or more, the method promotes in-vitro amplification of the NK cell, and the obtained NK cell has the advantages of being high in purity and large in number, saving the cost and improving the killing activity.

The invention relates to the technical field of cell in-vitro culture. The invention aims to provide an amplification method of high-purity anti-tumor NK cells. The amplification method comprises the following steps: S1, obtaining initial NK cells; s2, performing double-stage culture; a double-stage culture mode is adopted, the first-stage culture period is 8 days, the second-stage culture period is 10 days, and after the first-stage culture is finished, NK cells with full cytoplasm are separated through a density gradient centrifugation method for second-stage culture; s3, harvesting the NK cells. The method has the advantages of simplicity and convenience in operation, controllable cost, suitability for large-scale production and the like, and a more efficient and economical solution can be provided for preparation of the high-purity anti-tumor NK cells.

The application discloses a kind of multiple immunofluorescence detection method, equipment and medium based on space reconstruction, it is related to histopathological analysis technical field, including, preparation tissue sample and join fluorescent reference microbead to obtain continuous section;To the tissue sample cycle pretreatment, and utilize fluorescent reference microbead to carry out intensity normalization and bleaching time shift correction, obtain correction image sequence;Spectral unmixing is carried out under three-dimensional voxel coordinate system and introduces space regular constraint, obtain voxel level fluorescence intensity vector;Combining nuclear dye signal and fluorescent reference microbead, using rigid and non-rigid registration and supplemented with three-dimensional deconvolution, reconstruct high-resolution voxel stack and segment cell voxel set;The adjacent relationship and colocalization index of different phenotype cells are calculated, and detection result is generated.The application realizes high-resolution tissue and cell reconstruction under three-dimensional space, improves spatial continuity and accuracy, and improves the stability and accuracy of voxel level fluorescence signal quantification.

The invention discloses a functionalized culture medium, a culture medium composition and a culture method for NK cell amplification, and belongs to the field of biological medicine. Aiming at the problem that the activity and the purity of the NK cells cannot be considered at the same time in the existing NK cell in-vitro amplification, the invention provides a functionalized culture medium for NK cell amplification, and the functionalized culture medium comprises a proliferation culture medium and an activation adaptation culture medium; the proliferation culture medium and the activated adaptive culture medium both comprise IL-10, and the concentration of the IL-10 in the proliferation culture medium is lower than that of the IL-10 in the activated adaptive culture medium. IL-10 is added into a proliferation culture medium and an activation adaptation culture medium, the concentration of IL-10 in the two culture mediums is different, and the synergistic effect of high purity (larger than 95%) and high activity (the killing rate is larger than 75%) is achieved by inhibiting T cell proliferation and enhancing the anti-tumor microenvironment capacity of NK cells.

The invention discloses an NKT cell in-vitro amplification method based on a combined stimulant, and belongs to the technical field of biology. The method comprises the following steps: firstly separating mononuclear cells in autologous peripheral blood, and then performing induced activation and amplification on the NKT cells in a serum-free culture system by adding a group A streptococcus preparation, a zoledronic acid preparation and a cytokine IL-2 as a combined stimulant. The method is easy to operate, efficient amplification of the NKT cells can be achieved only through three key reagents, the obtained NKT cells have the advantages of being large in number, high in purity, high in cytotoxicity and the like, and the requirements of clinical treatment can be met. Compared with a traditional method, the culture cost is remarkably reduced, the operation process is simplified, large-scale production, popularization and application are easy, and an efficient and economical technical scheme is provided for NKT cellular immunotherapy.

The application relates to the technical field of cell in-vitro culture, and discloses a sphincter in-vitro culture environment construction device which comprises a culture device and a control host computer, the culture device is internally provided with a contraction member, the control host computer is connected with the culture device in a matched mode, and is used for regulating and controlling the action of the contraction member, so as to construct a physiological contraction environment. The application can simulate the annular contraction and the physiological contraction characteristics of the high and low pressure of the human sphincter, restore the dynamic contraction process, adopt a detachable structure, adapt to different culture cavities, has strong experimental flexibility, combines a perfusion design with a three-dimensional culture layer, simulates in-vivo nutrition transportation and metabolism discharge, provides a bionic growth environment for cells, integrates strain force detection and temperature control closed-loop regulation, is accurate in parameters and reliable in data, makes up for the short board of traditional static culture and animal experiments, provides a stable and efficient experimental platform for sphincter disease mechanism, mechanical response research and drug screening.

The present invention discloses a method and device for automatically identifying and segmenting cell somas based on two-photon calcium imaging data. First, a two-photon calcium imaging video of cells is acquired, and image denoising and image enhancement are performed on each frame of the video image. Then, the two-photon calcium imaging video is reduced in dimension to a locally correlated summary image, and a cell recognition algorithm based on multi-scale dot enhancement is applied to the locally correlated summary image to calculate the central coordinates of near-circular structures of different sizes within the imaging region, i.e., the seed points of the cell somas. Finally, for each seed point, the two-photon calcium imaging video is cropped into a block video of a fixed size centered on the seed point, and a cell segmentation algorithm based on an elliptical shape-constrained active contour model is applied to the block video to obtain the contour of the cell soma. The present invention can improve the operation efficiency and reduce the data storage volume.

The invention belongs to the technical field of biological medicine research, and particularly relates to a cell in-vitro calcification inducer, an induction culture medium and application thereof, an aortic valve calcification in-vitro induction model and a modeling method thereof. The inducer provided by the invention is prepared from the following components in parts by weight: 2160.4 parts of beta-sodium glycerophosphate, 0.039 parts of dexamethasone, 50 parts of vitamin C and 34 parts of vitamin D. The invention further provides a culture medium formed by using the inducer and a method for modeling an aortic valve calcification in-vitro induction model by using the inducer. According to the method, osteogenic differentiation of the aortic valve interstitial cells can be stably induced within a short time, an efficient and repeatable standardized induction scheme is provided for in-vitro aortic valve calcification models and drug screening, and the method has a very good application prospect.

The invention relates to application of lentinan in preparation of a medicine for enhancing Kukuzhou cell immune function, and belongs to the technical field of immunopharmacology. The invention discloses a mechanism and application of lentinan LNT (Lentinan) based on a TLR2 (Toll-Liver Receptor 2) signal channel for enhancing the immune function of liver cumness cells and promoting lysosome maturation. By constructing an LNT fluorescence labeling system, a Kuptake cell in-vivo tracing model and a TLR2 function intervention model, it is proved that LNT can be directly combined with TLR2 and activate a TLR2 / MyD88 / NF-kappa B signal axis, and therefore the phagocytosis and immune response capacity of Kuptake cells is enhanced. Meanwhile, TLR2 signal-induced reactive oxygen species (ROS) up-regulation can promote TFEB dephosphorylation and nuclear translocation, lysosome maturation is driven, and degradation metabolism of LNT is accelerated. The invention provides a TLR2-based immune enhancement mechanism of the LNT, and provides a theoretical basis and an application basis for developing immunomodulatory targeted drugs.

The present disclosure provides compositions, kits, and methods for promoting in vitro maturation of cells. The compositions, kits, and methods utilize at least one inhibitor of an epigenetic regulator and at least one agonist of a calcium channel. Suitable applications of the present disclosure include use in maturing and differentiation stem cells into viable mature neuronal cells for use in disease modeling, prevention and treatment. The present disclosure also provides methods of screening compounds that are suitable for promoting in vitro maturation of cells.

The invention provides application of mesenchymal stem cell-derived small extracellular vesicles in preparation of a product for preventing and / or treating airway inflammation mainly caused by neutrophil infiltration. Human cell in-vitro experiments and mouse in-vivo experiments find and prove that MSC-sEV can be directly taken by cells, activation of a cell JAK2-STAT3 pathway is inhibited, Th17 cell polarization is inhibited, neutrophil airway inflammation is remarkably relieved, and protein of MSC-sEV plays an important role in inhibition of Th17 cell polarization. The invention provides the application of the mesenchymal stem cell-derived small extracellular vesicles to prevention and treatment of airway inflammation mainly aiming at neutrophil infiltration, has the advantages of safety and effectiveness, and provides a new medicine for clinic.

The application relates to a G-quadruplex RNA fluorescent ligand and a preparation method and application thereof, a structure formula of the G-quadruplex RNA fluorescent ligand is shown in the following formula: the compound can be specifically targeted and can be subjected to fluorescent imaging in a solution system and a cell system, a high-throughput fluorescent screening method is established in the solution and the cell system, and an organic small-molecule compound with strong binding capacity to G-quadruplex RNA can be effectively identified and screened from a small-molecule drug library; the screening result of the method is high in accuracy, the affinity and biological activity of a candidate compound can be quickly determined by evaluating the change of a fluorescent signal, and therefore the research and development process of an antitumor drug is accelerated; and the application belongs to the field of biological medicines.

The invention provides an extraction method of high-performance gamma delta T immune cells and application of the high-performance gamma delta T immune cells in tumors. The method comprises the following steps: firstly, separating mononuclear cells from peripheral blood, carrying out activation induction by using a serum-free culture medium containing zoledronic acid, IL-15 and OKT3, and supplementing a culture medium containing IL-2 according to a specific proportion for multiplication culture from the third day of culture. Through the optimization process, the expression rate of TCR-gamma delta < + > is as high as 96.84%, and gamma delta T cells with high activity are obtained. An in-vivo tumor inhibition experiment shows that the cell has a remarkable growth inhibition effect on liver cancer nude mouse transplanted tumor, the effect is equivalent to that of a positive control drug, and obvious toxic and side effects are not observed. The method is stable in process and clear in component, and a reliable technical basis is provided for clinical transformation of gamma delta T cells in immunotherapy of solid tumors and hematologic tumors.

The invention belongs to the technical field of cell therapy, and relates to a serum-free and heterologous component-free NK cell culture medium as well as a preparation method and application thereof. The culture medium comprises a classical culture medium, a proliferation promoting component, vitamins, trace elements, lipids, transferrin, an energy substance and beta-mercaptoethanol, the proliferation promoting component is prepared from human albumin, resveratrol, insulin and hydroxyproline. On the basis of a serum-free and heterologous component-free NK cell culture medium, IL-12, rapamycin, polyinosinic-polycytidylic acid and human AB plasma are additionally added to obtain an activated culture medium, the activated culture medium and an amplification culture medium are jointly used, the culture effect is better, and amplification components IL-2 and human AB plasma are additionally added to obtain an amplification culture medium. Through precise component design, the NK cell in-vitro proliferation efficiency, motility, purity and killing ability are remarkably improved, and key problems existing in a traditional culture medium are efficiently solved.

The application discloses a kind of mixed immune cells and its culture system and in vitro culture method.The mixed immune cells include 75%~80% of γδT cell, 10%~15% of NK cell and 1~2% of NKT cell.The in vitro culture method of the application includes, when peripheral blood mononuclear cell is inoculated with activation culture medium, adding immune cell first activation component, adding immune cell reactivation component after 0~6 hours of inoculation, continuous activation culture;Then the activated culture cell is transferred into the coated culture plate or culture bottle, and the mixed immune cell is obtained by expansion culture.The mixed immune cell of the application has higher liver cancer cell HepG2 killing effect.The mixed immune cell in vitro culture method of the application has high expansion efficiency, and the expansion multiple can reach 2000-8000 times.

The invention discloses a method for confirming the cell mutation degree in a TCT detection mode. The method comprises the following steps: 1) preprocessing an original TCT cell staining image; 2) dividing pixels of the obtained grey-scale map into three categories of cell nucleuses, cell bodies and backgrounds, and respectively completing corresponding loop iterations; 3) performing segmentation, post-processing and closed operation in sequence from the obtained result maps of the three clusters, and determining a cell nucleus region result; 4) sequentially carrying out segmentation and post-processing operation in combination with the obtained cell nucleus region from the three obtained clustering result maps, and determining a cell body region result; and 5) according to the obtained cell nucleus region result and the obtained cell body region result, obtaining the nucleus-plasma ratio rho of the cells, and finally obtaining the characterization conclusion of the cell mutation degree. The method belongs to the technical field of images, and solves the problem that the cell mutation degree cannot be accurately determined due to the fact that the nucleus-plasma ratio of the cells is difficult to accurately calculate in the prior art.

The invention provides a culture medium and an in-vitro induction method for in-vitro erythroid induction of trace primary erythroid terminally differentiated cells, and belongs to the technical field of erythrocyte induction. The invention provides a culture medium for inducing primary erythroid terminal differentiated cells into erythrocytes in vitro. The culture medium comprises a first culture medium added with a plurality of inducing factors and a second culture medium not added with the inducing factors. The invention also develops a method for inducing micro primary erythroid terminal differentiated cells into erythrocytes in vitro, the method is suitable for primary cells, and the problem of drug response distortion caused by large difference between an immortalized cell line and cells in a living body is effectively avoided. According to the in-vitro induction method disclosed by the invention, the maturing and differentiating process can be completed in vitro only by using a very small amount of primary cells, so that the dependence on precious clinical samples is greatly reduced, and repeated experiments and high-throughput screening by using limited samples become possible.

The invention discloses a cell co-culture flow chamber device for simulating a blood brain barrier in vitro, and belongs to the technical field of cell in-vitro culture. The cell co-culture flow cavity device for simulating the blood brain barrier in vitro comprises a blood flow bottle and a peristaltic pump which are communicated into a loop through a connecting pipe, the peristaltic pump pumps liquid in the blood flow bottle into the connecting pipe to simulate blood flow, the cell co-culture flow cavity device comprises a semi-permeable membrane pipe which is arranged on the connecting pipe at the outlet end of the peristaltic pump, and the connecting pipe is provided with a broken opening; two ends of the broken opening are respectively communicated with two ends of the semi-permeable membrane tube; culture medium liquid and experimental cells are injected into the first culture dish, and the two opposite side walls of the first culture dish are connected with the connecting pipes on the two sides of the broken opening respectively, so that the semi-permeable membrane pipe is located in the first culture dish. The cell co-culture flow chamber device for simulating the blood brain barrier in vitro avoids the problems that ethical problems exist and experimental conditions are difficult to control in the process of drug research on BBB at present, and is simple to operate and obvious in experimental effect.

The present invention relates to a PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab / IPH4301 nanoprobe and an application thereof. The prepared nanoprobe is characterized, the binding ability of the nanomaterial and NK-92MI cells is analyzed by flow cytometry, MRI is used to image the NK-92MI cells labeled with the nanoprobe in vitro and the T1-weighted (T1WI) signal changes are analyzed, Western blot is used to detect the expression of apoptosis-related proteins in breast cancer MDA-MB-231 cells after co-culture, and Elisa The expression level of IFN-γ in the culture system was detected by the PEG-G5.NH2-FITC-DOTA(Gd)-Monalizumab / IPH4301 nanoprobe of the present invention was found to be able to bind to NK-92MI cells and to be imaged in vitro using 3.0TMRI. After being modified with Monalizumab and IPH4301 double antibodies, the nanoprobe promoted the release of IFN-γ by NK-92MI cells and enhanced its ability to kill MDA-MB-231 cells, providing an experimental basis for in vivo imaging and immunotherapy.

The invention discloses a dry memory T cell in-vitro induction and amplification method based on metal ion regulation and application thereof. According to the method, Mn < 2 + > is added in the cell culture process to effectively induce the T cells to differentiate to the dry memory phenotype, so that the production cost of the dry memory T cells is greatly reduced while the use of other cell factors and chemical inducers is reduced. The dry memory T cell obtained by adopting the method has good multiplication capacity and functional activity, is suitable for clinical treatment, and particularly shows a wide application prospect in anti-tumor immunotherapy.

Methods of treating diseases associated with pathologic neuronal cells are provided. Accordingly, there is provided a method of treating a disease associated with pathologic neuronal cells characterized by aberrant excitability comprising: (a) administering into a first neural region of the subject a therapeutically effective amount of a polynucleotide encoding a bistable type II opsin attached to a heterologous ER export signal and / or membrane trafficking signal which enables trafficking of said bistable type II opsin to axon and / or dendrite terminals, wherein said first neural region comprises cell bodies of said pathologic neuronal cells; and (b) exposing a second neural region of said subject to light in a wavelength that activates said bistable type II opsin, wherein said second neural region comprises axon or dendrite terminals of said pathologic neuronal cells associated with a symptom of said disease, and wherein said first neural region and said second neural region are distinct.

The present invention relates to a method for mass proliferation and culture of iNKT cells, iNKT cells produced by the method, and uses thereof, the method comprising the steps of: expanding iNKT cells from peripheral blood mononuclear cells; and mass-proliferating and culturing the iNKT cells by treating same with an anti-CD28 antibody, the CD40 protein, and an anti-TCR Vα24-Jα18 antibody. Since the method for mass proliferation and culture of the present invention can produce iNKT cells while maintaining an excellent proliferation rate and purity without artificial antigen-presenting cells, the method can be effectively utilized in related technical fields such as treatment of cancer or inflammatory diseases using iNKT cells.

The application discloses a Turn-Off type cell in-vivo tracing probe and a preparation method and application thereof, and the probe structure is composed of three parts, including a membrane anchoring part, an enzyme specificity response part and a fluorescent group. The probe can mark cells through the hydrophobic tail chain of the membrane anchoring part, at this time, the fluorescent group part emits near-infrared fluorescence, and shows a Turn state. When the adoptive cells are apoptotic in the body, the probe falls off from the cells. The free probe falling off is phagocytosed by macrophages in the body, in the intracellular macrophages, the enzyme specificity response part in the probe structure responds to enzyme bond breaking, the fluorescent group is dissociated, does not emit light, and shows an Off state. Therefore, the fluorescent probe can not only trace the in-vivo living cells and monitor the death and life of the cells in the body, but also can effectively overcome the fluorescent "pollution" phenomenon caused by the persistent existence of the fluorescent of the free probe falling off, and provides a new visual tool for monitoring the in-vivo fate of the living cell drugs.

The application discloses application of ostruthin in enhancing CAR-T cell anti-tumor function, belongs to the technical field of biological medicine, and the ostruthin can promote cell proliferation, inhibit cell exhaustion, promote the secretion of cytokines related to killing cancer cells, and further enhance the anti-tumor function of CAR-T cells in vivo and in vitro as a cell enhancer of CAR-T cells; experimental results show that when ostruthin is combined with CAR-T cells for anti-tumor treatment, compared with ostruthin or CAR-T cells alone, the combination of the two can significantly inhibit the growth of solid tumors. The application provides a new technical means for improving the tumor treatment effect of CAR-T cell immunotherapy.

The application provides a method for enhancing the expansion and activation of memory-like NK cells, and belongs to the technical field of cell culture. The application provides a method for obtaining strong memory-like NK cells, which comprises using complete culture medium, IL-2, IL-12, IL-15, IL-18, sodium alpha-ketoglutarate and DOT1L inhibitor; the concentration of the sodium alpha-ketoglutarate is 0.1-2 mM, and the concentration of the DOT1L inhibitor is 10-100 nM. The addition of low-concentration sodium alpha-ketoglutarate in the complete culture medium of memory-like NK cells can promote the in-vivo and in-vitro proliferation capacity of the memory-like NK cells. The addition of low-concentration DOT1L inhibitor in the late culture stage can further enhance the killing function of the memory-like NK cells by inhibiting H3K79me2 / 3, etc., so that memory-like NK cells with a larger number and stronger killing capacity are obtained.

The application discloses a culture medium and application thereof, and a method for in-vitro separation and culture of goat intramuscular precursor fat cells and adipogenic differentiation of the cells, and relates to the technical field of cell culture. The culture medium comprises the following components: 8-12 mu g / mL insulin, 8-12% fetal bovine serum in volume fraction, 95-105 U / mL penicillin, 95-105 mu g / mL streptomycin, and the like. On the basis of the prior art, the application mainly adds appropriate glycyrrhizin and pilocarpine in the culture medium, and finds that the two components are in cooperation, and the adipogenic differentiation ability of cells such as mesenchymal stem cells and precursor fat cells is significantly improved, and the application range is wide. On the basis, the application further provides a complete method for in-vitro separation and culture of goat intramuscular precursor fat cells and adipogenic differentiation of the cells, and provides a method and a way for a series of researches on goat primary intramuscular precursor fat cells in the future.

This invention discloses the application of T cells overexpressing PRDM16 in the preparation of anti-tumor or infectious disease drugs. Through experiments, this invention verifies that overexpression of PRDM16 enhances the in vivo and in vitro activity of T cells. It was found that in vitro overexpression of PRDM16 significantly enhances the killing effect of T cells on target cells. In vivo reinfusion of PRDM16 has stronger anti-tumor and anti-infection functions compared to wild-type T cells. Therefore, it can be used in malignant tumors and infectious diseases to enhance the efficacy of T cell adoptive therapy and solve the defect of T cell adoptive therapy where the killing power of target cells is suppressed due to environmental influences.