45 results about "Cell phenotype" patented technology

The present application relates to the field of bioinformatics. In particular, the present application relates to methods and apparatus for speculating variable splicing functionality based on single cell transcriptome data. The method comprises the following steps: determining a variable splicing mode of each gene in a data set in a cell; determining the incidence relation between the variable splicing mode and the gene expression of each gene; a variable splicing mode module is determined according to the incidence relation between the variable splicing modes and the gene expression, and the variable splicing mode module is a variable splicing mode set obtained through clustering according to the correlation between the variable splicing modes and the cell phenotypes; displaying the cell splicing heterogeneity according to the variable splicing mode module; and / or determining a potential regulatory mechanism between the variable splicing mode and the gene expression according to the variable splicing mode module, the potential regulatory mechanism being used for embodying key splicing factors in the gene expression, and a biological approach in which the variable splicing mode affects the cell phenotype.

The present invention relates generally to compositions, systems and methods for modulating hepatitis B virus (HBV) by targeting gene repression. In particular, the present invention provides an epigenetically modified DNA targeting system, such as a CRISPR-Cas / guide RNA (gRNA) system, for transcriptional repression of hepatitis B virus genes to promote cell phenotypes that lead to reduced HBV infection. In some embodiments, the epigenetically modified DNA targeting systems of the present invention bind to or target a target site of at least one gene in the hepatitis B virus DNA sequence or a regulatory element thereof in a cell. In some aspects, the systems provided herein relate to transcriptional repression of one or more hepatitis B virus genes and / or regulatory elements thereof. In some aspects, the invention also provides methods and uses associated with the provided compositions, such as in repression of hepatitis B virus replication and expression associated with hepatitis B infection.

A composition is provided comprising a cell population and pharmaceuticals derived from human gingival fibroblasts, present in proportions not found in natural gingival tissue, but having a cellular phenotype preferentially selected to express proteins favorable for angiogenesis and anti-inflammatory effects, while reducing the formation of cell populations that promote tumorigenesis and / or metalloproteinases that inhibit tissue regeneration. A method for producing such a composition that increases proliferation several times over compared to previously known methods is provided, and a method for using such a composition in a wide range of human cell therapies is also provided.

The invention provides an application and a method for inducing phenotype transformation from oral mucosa epithelium to dry lip epithelium by linoleic acid, linoleic acid with proper concentration acts on oral mucosa epithelial cells to regulate and control cell differentiation procedures, so that expression of specific markers of the oral mucosa epithelial cells is reduced, expression of specific markers of dry lip epithelial cells is increased, and expression of the specific markers of the oral mucosa epithelial cells is increased. The directional transformation of the cell phenotype is realized. The linoleic acid with the proper concentration is 10-20 [mu] M. The linoleic acid has the effects of regulating keratinization index expression of mucous membrane epithelial cells and enhancing the mucous membrane tissue barrier function for the first time, and the function is not recognized in previous research and clinical application of the linoleic acid.

The invention discloses application of ACSS2 as an action target in screening or preparing a medicine for preventing or treating virus-induced vascular remodeling diseases, and belongs to the field of biological medicine. It is found that by inhibiting ACSS2 gene expression or protein activity, Zika virus induced vascular smooth muscle cell phenotypic transformation can be remarkably reversed, and occurrence and development of angiotensin II / aminopropionitrile induced aortic aneurysm / dissection are relieved. The invention provides a siRNA sequence (SEQ ID NO.1-2) specifically targeting ACSS2 and a pharmaceutical composition containing the siRNA, and provides a novel treatment target and a precise intervention tool for virus-related vascular remodeling diseases.

The invention relates to a drug effect evaluation method for fusing target molecules and time-concentration dependent cell phenotypes, which comprises the following steps: S1, preparing samples including a modeling sample and a to-be-detected sample; s2, carrying out FRET imaging and cell fluorescence imaging; s3, carrying out FRET image processing and FRET efficiency calculation; s4, cell fluorescence image processing and feature extraction; s5, calculating a phenotype characterization value; s6, calculating an FRET characterization value; s7, basic drug response value calculation; and S8, comprehensive drug effect evaluation. Target molecule information and time-concentration dependent cell phenotypic response are fused, a comprehensive drug effect evaluation model is constructed, the effect of drugs on whole cells is reflected, and whether the drugs target specified molecules or not is specifically indicated.

The invention relates to the technical field of biomedical processing, and particularly provides a cryopreservation method of cord blood mononuclear cells, which comprises six steps of whole blood separation, plasma inactivation, CBMC cell separation, cryopreservation liquid preparation, cell cryopreservation and detection after cryopreservation recovery. The CBMC cell separation comprises three sub-steps; performing cell cryopreservation, namely averagely dividing the separated CBMC into a plurality of parts, filling the CBMC into a cryopreservation tube according to a certain density, putting the cryopreservation tube into a programmed cooling instrument, and performing cryopreservation according to a plurality of programs; and detecting after cryopreservation and resuscitation, wherein the detection comprises three sub-steps of cell viability detection, cell recovery rate detection and cell function detection. The problems that in the cryopreservation process of a traditional method, cell viability is reduced, the number of cells is greatly reduced, cell adhesion and caking are caused, cell functions are abnormal, cell phenotypes are changed after recovery, functional activity is damaged and the like are solved, the utilization rate of the CBMC is increased, and the technical scheme is obviously superior to the traditional cell cryopreservation method; the viability, the recovery rate and the function of the recovered cells are obviously improved.

Techniques for identifying a cellular phenotype indicative of a condition of a subject are described. The techniques can include identifying sequence read data indicating sequences of DNA fragments from a sample from a subject; determining, based on the sequence read data, endpoint positions of the DNA fragments; determining input features based on the endpoint positions of the DNA fragments; and predicting, using a classifier and based on the input features, a cellular phenotype indicative of a condition, such as a cancer, within the subject.

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.

Some aspects relate to engineered cells comprising a nucleotide sequence encoding a chimeric antigen receptor (CAR) and at least at first coding exon of a FOXP3 gene, where expression of the CAR and FOXP3 are linked, such that CAR+ cells have a regulatory T cell (Treg) phenotype. Some aspects relate to methods of producing engineered cells in which expression of a CAR and FOXP3 are linked, thereby reducing the incidence of contaminating CAR+ T effector cells. Further aspects relate to methods of screening CARs suitable for expression by a Treg. Some aspects relate to engineered cells expressing an anti-CD19 chimeric antigen receptor (CAR). Some aspects relate to methods of producing engineered cells expressing an anti-CD19 CAR. Some aspects relate to uses of cells expressing anti- CD19 CARs.

The application discloses immortalized chicken embryo epithelioid cells and a preparation method and application thereof, and the preparation method of the immortalized chicken embryo epithelioid cells comprises the following steps: extracting primary chicken embryo epithelial cells from a chicken embryo; and culturing the primary chicken embryo epithelioid cells in a conditional medium containing chicken Noggin protein for subculture, so as to obtain the immortalized chicken embryo epithelioid cells. The immortalized chicken embryo epithelioid cells prepared by the preparation method can be stably subcultured for a long time, can maintain stable proliferation ability after multiple subcultures, reduces individual differences, and improves the reliability and repeatability of experimental results. In addition, the preparation method does not introduce exogenous genes or change the genotype of the cells, ensures the stability at the gene level, and avoids the interference of the exogenous genes on the stability of the cell phenotype.

The present application relates to the use of polysialic acid or its derivatives in anti-foreign body reaction, in particular, to a polysialic acid anti-foreign body reaction material for resisting biological contamination, for regulating macrophage phenotype to reduce inflammatory response, and for resisting foreign body reaction. The polysialic acid material includes a bulk material prepared from polysialic acid and its derivatives, or a coating material for surface modification of the material, which can resist protein adsorption, cell adhesion and adhesion of various microorganisms, can act on immune cells, such as regulating the phenotype of macrophages, thereby reducing acute and chronic inflammatory response, and has excellent anti-foreign body reaction effect as an implant material.

The invention relates to the technical field of medicines, in particular to a zinc ion polypeptide compound immunologic adjuvant as well as a preparation method and application thereof. The preparation method comprises the following steps: mixing polypeptide, a PBS buffer solution, new coronavirus S protein, a CpG ODN immune activator and zinc ions, and standing to obtain the zinc ion polypeptide compound immunologic adjuvant. A dual-adjuvant system mixed with CpG ODN shows excellent performance in activation of lung tissue immunity, and in the cellular immunity level, the response intensity of CD4 + T and CD8 + T cells is enhanced, and formation of related tissue resident memory T cell phenotypes is promoted; on the humoral immunity level, a Gel (Zn < 2 + >) and CpG ODN dual-adjuvant system obviously stimulates the increase of the number of B cells and the activation, and can effectively stimulate the secretion of a main antibody sIgA for mucosal immunity. On the whole, the dual adjuvants realize the synergistic enhancement of T cell (containing TRM) and B cell immunity.

A method for predicting drug pathways that modulate cellular phenotypes is provided. The method includes obtaining a logic-based differential equation model of a signaling network relating to a cellular phenotype, comprising nodes representing proteins or mRNAs and edges representing interactions. The method further includes identifying candidate compounds affecting the cellular phenotype, determining protein targets using a drug-target database, and mapping pathways from protein targets to network nodes by searching a protein interaction database and computing ranked pathways by minimizing a cost function based on edge weights. The method includes expanding the model by incorporating mapped pathways, simulating expanded model variants to predict compound effects on the cellular phenotype, and outputting predictions of how candidate compounds modulate the cellular phenotype through identified pathways.

The present disclosure provides a method for screening cells, the method including a step of preparing a plurality of cells which are tagged with a first barcode nucleic acid associated with a test target and treated with the test target, a step of sorting the plurality of cells based on cellular phenotype using an imaging cell sorter, and a step of identifying the test target used to treat each cell using the first barcode nucleic acid as an indicator.

Processes to characterize classes and / or phenotypes are provided. In some instances, omic data is used to infer a class status or phenotype. Biomarkers for many potency phenotypes are also described.

The application discloses a detection system and method for CAR-T cell phenotype analysis and function quality, which comprises nucleic acid aptamer, including at least one of CD3 aptamer, CD25 aptamer and CD122 aptamer; nucleic acid nano framework; wherein the nucleic acid aptamer is combined with the nucleic acid nano framework, and can produce a fluorescent signal when the CAR-T cell is activated. The detection system combines nucleic acid aptamers for CD3, CD25 and CD122, and can quickly, without washing, sensitively analyze the phenotype of the CAR-T cell and control the function quality. The detection system utilizes the programmability of the nucleic acid nano structure, realizes real-time monitoring of the activation state and cytotoxicity of the CAR-T cell, has the characteristics of high specificity, wide dynamic range and strong serum stability, can distinguish the resting and activated T cells within 30 minutes, and can capture the phenotype and function difference of the CAR-T cell in a co-culture experiment.

The invention discloses an application of a gene for coding LRP1-beta chain oligopeptide as a target in screening / preparing a medicine for treating aortic dissection, an application of a reagent for inducing overexpression of the LRP1-beta chain oligopeptide in preparing the medicine for treating the aortic dissection, and an application of the LRP1-beta chain oligopeptide as a biomarker in preparing an aortic dissection diagnostic reagent. Experiments prove for the first time that the LRP1-beta chain oligopeptide can inhibit phenotypic transformation of vascular smooth muscle cells and inflammatory infiltration of macrophages by down-regulating expression of OPN, alleviate inflammatory response and relieve or inhibit occurrence and development of aortic dissection, provides a new direction and thought for treatment of aortic dissection, and has broad application prospects in treatment of aortic dissection. The compound is expected to be applied to research and development of new drugs for treating aortic dissection

Provided are epigenetic-modifying DNA-targeting systems, such as CRISPR-Cas / guide RNA (gRNA) systems, that bind to or target a target site in a gene or regulatory element thereof in a T cell. In some aspects, the provided epigenetic modifying DNA-targeting systems modulate a T cell function, such as a T cell phenotype or activity. In some aspects, also provided herein are methods and uses related to the provided compositions, for example in modulating T cells including in connection with methods of adoptive T cell therapy.

The invention discloses application of flueggeine B or pharmaceutically acceptable salt thereof in preparation of a medicine for treating pulmonary fibrosis. The chemical structure of the flueggeine B is shown as a formula (I). The flueggemine B shows an improvement effect in a bleomycin-induced pulmonary fibrosis mouse model, improves the weight and lung function parameters of a mouse, relieves the degree of pulmonary fibrosis, has an inhibition effect on lung tissue collagen deposition, can also effectively improve the phenotype of pulmonary epithelial cells and inhibit abnormal activation of myofibroblasts, and can be used for treating pulmonary fibrosis. Therefore, the traditional Chinese medicine composition has a remarkable relieving effect on bleomycin-induced pulmonary fibrosis.

The invention discloses a microcarrier for in-vitro amplification of mesenchymal stem cells. The microcarrier comprises a microsphere matrix formed by a biocompatible synthetic polymer, a nitrogen-containing functional group layer formed on the surface of the microsphere matrix, and recombinant human RGD-Fc fusion protein covalently linked to the nitrogen-containing functional group layer. Wherein the nitrogen-containing functional group layer is obtained through ammonia plasma treatment, the recombinant human RGD-Fc fusion protein contains a human RGD peptide fragment and a human immunoglobulin Fc structural domain, and the microcarrier does not contain any animal source component. The microcarrier can support stable amplification of mesenchymal stem cells to the eighth generation or above in a serum-free and animal-source-free culture system, and the cell phenotype positive rate is not lower than 90%. The invention solves the problems of heterologous risk, batch instability and non-specific adhesion of the existing animal-derived microcarrier, and is suitable for large-scale production of clinical-grade cell therapy products.

The invention relates to the technical field of stem cells, in particular to a preparation method of human placenta amnion pluripotent stem cells. The method specifically comprises the following steps: S1, tissue pretreatment and physical pre-layering; s2, carrying out mechanical-enzymatic synergistic dynamic dissociation; s3, gradient purification and washing; s4, performing soft matrix phenotype maintenance culture; according to the method disclosed by the invention, the separation efficiency of amniotic epithelial cells is improved and the enzyme exposure load is reduced through synergistic enzymolysis of Dispase II and Trypsin in combination with weak shearing force generated by the magnetic beads. Meanwhile, SB431542 blocks a signal channel to prevent EMT, a GelMA soft matrix maintains cell phenotypes, and Percoll density gradient centrifugation is matched to improve cell purity.

The present disclosure is related to systems and methods to be used as aids in the diagnosis of risk for oral cancer by identifying cellular phenotype characteristics of a noninvasive brush biopsy sample.

The present application relates to the biomedical field, and particularly relates to a gene modified cell preparation device and application thereof. The present application provides a device for bedside preparation of gene modified immune cells, wherein a gene modification system can complete gene modification of T cells in a short time, which can not only realize faster preparation of gene modified T cells, but also has better original cell phenotype due to short-time gene modification of the prepared gene modified T cells, and has better in-vivo persistence and safety, thereby realizing ultra-short-time gene modification of resting T cells, and providing a possibility for bedside CAR-T cell preparation.

The application discloses a ctPistol ribozyme and application thereof in gene silencing, and belongs to the technical field of biology.The disclosed ctPistol ribozyme can realize effective control of cell phenotype by silencing the expression of key genes.In addition, when applied in a mouse cancer model, the ctPistol realizes a powerful and persistent anti-tumor effect by targeting oncogenic driver genes.As a powerful gene silencing tool and a promising RNA-based therapeutic candidate drug, the ctPistol ribozyme shows an effective way of developing diversified ribozyme-mediated gene control technology.

A biosensor comprises a complementary metal-oxide semiconductor (CMOS) image sensor (CIS), wherein the CIS comprises a plurality of light sensitive diodes disposed over a silicon substrate, and one or more metal layers for interconnect disposed therein above, and wherein the silicon substrate is back-lapped to a predetermined thickness. It further comprises a microwell layer disposed over the back-lapped silicon layer having etched therein an array of microwells, each microwell having a bottom which is the back-lapped silicon substrate and is further adapted to capture therein one or more biological cells. The biosensor can be used to train an artificial intelligence (AI) engine to generate an AI model for inference or prediction of a phenotype of a biological cell captured by the biosensor. The training is performed on images from both a fluorescence microscope and the biosensor.

Provided herein are epigenetic-modifying DNA-targeting systems, such as CRISPR-Cas / guide RNA (gRNA) systems, for the transcriptional repression of Hepatitis B viral (HBV) genes to promote a cellular phenotype that leads to the reduction of HBV infection. In some embodiments, the epigenetic-modifying DNA-targeting systems bind to or target a target site of at least one gene or regulatory element thereof in a Hepatitis B viral DNA sequence in cell. In some aspects, the provided systems relate to the transcriptional repression of one or more Hepatitis B viral gene and / or regulatory element thereof. In some aspects, also provided herein are methods and uses related to the provided compositions, for example in repressing Hepatitis B viral replication and expression in connection with Hepatitis B infections.

The present application belongs to the field of cell culture. The present application provides a method for inducing, culturing and amplifying human natural killer cells, comprising the following steps: extracting peripheral blood from a healthy person, and separating PBMC; coating a container with an NK activation solution; culturing with an NK cell proliferation medium, inoculating PBMC, and incubating at a constant temperature for 1-6 days; and proliferating with an NK cell amplification medium, and incubating at a constant temperature for 1-10 days. The present application effectively improves the in-vitro proliferation rate of human natural killer cells. Through flow cytometry analysis, the proportion of NK cells with phenotypes of CD3, CD16 and CD56 in the cell population is all above 80%, and the cell phenotype is CD56 + CD3 ‑ NK cell group, and the positive rate is 97.57%. The present application has a simple composition formula, ideal culture effect, and wide application prospect.

The invention provides a method and system for constructing a metabolite and phenotype relation model and a storage medium, and the method comprises the steps: obtaining original cell morphological characteristic data, carrying out the preprocessing and batch effect correction of the original cell morphological characteristic data, and obtaining corrected cell morphological characteristic data; carrying out dimension reduction processing and clustering analysis on the corrected cell morphological characteristic data to obtain a plurality of cell phenotype clusters; constructing a reference space, projecting all cell data in the corrected cell morphological characteristic data to the reference space to obtain a projection result, and calculating a lever score of each cell based on the projection result; according to the cell phenotype clusters and the lever scores, constructing an incidence relation map between the metabolites and the cell phenotype clusters; and constructing an interpretable metabolite and phenotype prediction model based on the cell phenotype cluster, the corrected cell morphological characteristic data and the incidence relation map. According to the method, the specific influence of the metabolite on the cells can be accurately analyzed, and the reliable and predictable metabolite and phenotype relation model can be constructed.

The invention relates to the technical field of cytobiology and genes, in particular to an amniotic cell immortalized line establishment method based on a lentiviral vector, which comprises the following steps: acquiring primary cells from amniotic tissues, and removing fibroblast-like cells by a differential adherent culture technology to obtain a high-purity epithelial amniotic cell population. An immortalized gene is introduced into the purified cell by using the constructed lentiviral vector, and a plurality of monoclonal cell strains are established through drug screening and monoclonal separation. By detecting integration sites and expression levels of immortalized genes in all monoclonal strains, candidate cell strains with safe integration sites and stable expression are screened out, and finally the immortalized cell line is determined through passage and functional verification. According to the method, cell source purification and genetic integration evaluation are controlled, an immortalized amniotic epithelial cell line which is consistent in cell phenotype, safe in genetic background and stable in character can be established, and a reliable tool is provided for related research.