89 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.

An imaging flow cytometry-based high-throughput drug screening method, comprising: cell incubation, cell staining, acquiring single-cell images by using a flow cytometer, and image extraction and analysis, which combines the high-throughput advantages of flow cytometry and the imaging capability of a microscope, so that multi-channel single-cell images can be generated in a high throughput manner, thereby implementing acquisition of fluorescent images and unmarked images of single cells at a throughput of 102-105 cells per second, which can be used for cell phenotype drug screening and improving the screening efficiency by a factor of 102-104.

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 present disclosure relates, at least in part, to compositions comprising microgels and / or granular hydrogels with tailored surface biochemical properties that can serve, e.g., as artificial antigen-presenting cells (aAPCs) to mediate, e.g., T cell activation and expansion. Such microgels and / or granular hydrogels can be used to manufacture T cells for adoptive therapy in cancer treatment and tissue regeneration, and can be configured to serve, e.g., as a readily tunable and modular system to enable both rapid T cell expansion and control over T cell phenotype. Some aspects of the present disclosure provide methods and compositions for fabricating microgels and / or granular hydrogels, and modulating the surface properties of microgels and / or granular hydrogels by functionalizing, e.g., via layer- by-layer coating.

The present application provides a method for screening cells, the method comprising: a step of preparing a plurality of cells, the plurality of cells labeled with a first barcode nucleic acid and treated with a test target, the first barcode nucleic acid associated with the test target; a step of selecting the plurality of cells based on a cell phenotype using an imaging cytometer; and, a step of identifying the test target used to treat each cell using the first barcode nucleic acid as an indicator.

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 invention provides a composition of matter comprising a population of cells and an agent, which is derived from human gingival fibroblasts and has a cell phenotype present in a proportion that is not found in the natural gingival tissue, the cell phenotype being preferentially selected to express a protein beneficial to angiogenesis and anti-inflammatory action, meanwhile, cell populations of metalloproteinase formation that promotes tumorigenicity and / or inhibits tissue regeneration are reduced. The present invention provides methods of producing the compositions that can achieve multiple proliferation improvements compared to previously known methods, and methods of using the compositions in a wide range of human cell therapies.

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 present application relates to the application of delta-Viniferin in the preparation of drugs for preventing or treating metabolic cardiovascular diseases. Delta-Viniferin can inhibit the expression level of aging and inflammation related genes / proteins of vascular endothelial cells, reduce endothelial dysfunction, and clarify its anti-hypertensive effect. Delta-Viniferin can inhibit the expression level of vascular smooth muscle cell phenotype conversion related genes / proteins, improve the contraction function of vascular smooth muscle, and significantly alleviate the degradation of elastic fibers of the vascular wall and the rupture of the intima of the aortic wall in vivo, and clarify its effect on preventing and treating aortic dissection. Delta-Viniferin can inhibit the expression level of myocardial fibroblast fibrosis related genes / proteins, and clarify its anti-myocardial fibrosis effect. Delta-Viniferin can promote white adipose tissue browning by targeting the SIRT1-FOXO1 axis, increase energy consumption, and clarify its effect on resisting obesity. In summary, the compound delta-Viniferin has the effect of protecting various metabolic cardiovascular diseases, and can be used to prepare drugs for preventing and treating metabolic cardiovascular diseases.

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.

The present invention relates to a composition for improving the efficiency of cell suspension culture and a composition for improving the efficiency of cell adhesion culture. According to the present invention, it is possible to artificially alter the anchorage dependency of target cells to be cultured and to simply revert the altered phenotype, if necessary. According to the present invention, it is possible to alter the cell phenotype to an optimal state suitable for the culture purpose and environment and revert the altered phenotype to the original phenotype at a desired time point, thereby maximizing the efficiency of culture of various target cells, including host cells for recombinant protein production, as well as therapeutic immune cells and stem cells.

The invention discloses a method for preparing a dendritic cell vaccine through in-vitro amplification and induction of hematopoietic stem cells, and belongs to the field of biomedicine.The amplification induction method specifically comprises the following steps that firstly, peripheral blood is collected, the collected peripheral blood is diluted, and then peripheral blood mononuclear cells are separated; according to the invention, the conditions that cells enter an inflammatory emergency state due to a large dose of IL-4 in a traditional scheme, metabolic routes such as cell glycolysis are highly activated, ROS release is caused by oxidative stress, and cell dysfunction is caused by abnormal metabolism are avoided, and a cell combination closest to real DC cell phenotype and function in a human body can be obtained; the antigen presentation of the DC vaccine is improved to the maximum extent, meanwhile, hematopoietic stem cells can be partially differentiated to a myeloid system, DC progenitor cells and DC precursor cells, effective amplification of dozens of times can be achieved in quantity, and the technical bottleneck that the clinical curative effect is limited due to insufficient cell quantity is solved.

Provided herein are methods for determining a cell phenotype, such as an activation state, of a population of T cells. In some aspects, the provided methods are label-free methods. In some aspects, the provided methods can be used to monitor cell phenotypes, such as activation status, of T cell cultures. A computing device for use in performing the provided method is also provided herein.

The invention discloses application of a deubiquitinating enzyme STAMBPL1 as a pancreatic cancer diagnostic kit and in targeted drug development. According to the research, by mining a TCGA-PAAD public data set of a TCGA database, the transcriptome expression level of STAMBPL1 in pancreatic cancer tissue is found to be remarkably higher than that in normal tissue. The research also finds that the proliferation and migration of the human pancreatic cancer cell line PANC-1 can be inhibited in vitro by knocking down the STAMBPL1 through the small interfering RNA, and the mRNA level expression quantity of tumor proliferation and cell cycle related genes is reduced. According to the expression difference of the STAMBPL1 in pancreatic cancer derived from a patient and normal tissue and the experimental results of cell phenotype and gene transcription level expression detection after knockdown, the STAMBPL1 can be possibly used as a potential biomarker of pancreatic cancer and a potential target of a pancreatic cancer diagnostic kit.

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.

Inventors have synthesized peptides allowing an engagement of intact CD31 molecules on all the healthy endothelial cells and resting blood platelets and leukocytes that can enter in contact with an implanted device. Those cells can therefore receive the “leave-me-alone” signal delivered by the trans-homophilic engagement of CD31, which is essential to maintain the homeostasis in the circulation and vascularized tissues. Thrombotic or life-threatening occurrence of hemorrhagic or thromboembolic complications have impaired the use of endovascular devices. The devices bearing the mimicking peptides of the present invention are rapidly integrated, because they are perceived by blood platelets and leukocytes as a healthy endothelium, a “self” component. Furthermore, their ability to be rapidly endothelialized with a physiologic endothelial cell phenotype also limits platelet and leukocyte activation at the site of device implantation in the long-term. Accordingly, the present invention relates to peptides mimicking the trans-homophilic CD31-CD31 domain 1 and 2 intercellular interaction.

The invention relates to the technical field of cell therapy, and particularly discloses a repairable fibroblast preparation for treating pelvic organ prolapse. The active ingredients of the preparation are fibroblasts which highly express HAS1, POLR3G and DDR2 at the same time and do not express alpha-smooth muscle actin, and the preparation is compatible with pharmaceutic adjuvants such as normal saline, human serum albumin and dimethyl sulfoxide. The dosage form of the composition comprises an injection and a gel containing a hyaluronic acid matrix. The preparation method comprises the following steps: performing enzymolysis treatment to obtain primary cells, promoting repair phenotypic differentiation through estrogen induction, purifying target cells by adopting a fluorescence labeling flow sorting technology, and performing amplification in a serum-free culture medium to prepare a final preparation. According to the preparation, through the synergistic effect of fibroblasts and pericytes and the perivascular positioning characteristic, targeted repair of pelvic floor supporting tissue can be achieved; according to the preparation method, the cell phenotype stability and the preparation reproducibility are ensured by optimizing process parameters and quality control standards.

Disclosed is a method for in vitro expansion of tumor-infiltrating lymphocytes. Developed are a new method for isolating and enriching TIL cells in tumor tissues and an optimized culture method for TIL cells independent of feeder cells. 1010 grade or more of TIL cells having an anti-tumor function can be obtained. By means of cell phenotype analysis, compared with TILs expanded by the traditional two-step method, the cultured TIL cells have a higher proportion of memory T cells and a lower proportion of exhausted T cells and regulatory T cells.

The application relates to the technical field of culture medium prediction, and discloses a serum-free culture medium component prediction method based on artificial intelligence. The method comprises the following steps: constructing a phenotype condition vector according to a cell type identifier and a cell phenotype specification parameter, constructing a phenotype-formulation pairing data set according to historical culture batch data, training a conditional variational autoencoder with the phenotype condition vector as a conditional constraint, sampling a latent variable from a standard normal distribution, decoding the latent variable through a decoder to obtain a candidate culture medium formulation set, screening a candidate formulation with the minimum Euclidean distance through a forward verifier, and obtaining a recommended culture medium formulation. The application solves the problem that an existing serum-free culture medium component prediction method based on artificial intelligence cannot generate a culture medium formulation in reverse according to a clinical phenotype quality specification because the number of cells is taken as an optimization target.