135 results about "CD34" patented technology

The invention belongs to the technical field of biotechnology and animal models, and discloses a double-person-derived mouse model for simulating a tumor immune microenvironment and a construction method and application thereof. The method comprises the following steps: firstly, pretreating NSG immunodeficient mice by adopting low-dose whole-body irradiation in combination with double-antibody targeted bone marrow depletion, and transplanting CD34 + hematopoietic stem cells from the same human donor to complete human immune system reconstruction; separating tumor primary cells, tumor-related fibroblasts and tumor vascular endothelial cells of the same donor, performing three-dimensional co-culture to obtain homologous human tumor organs, and performing in-situ inoculation to immune reconstruction mice to obtain a target model. The core defects of MHC mismatching, low immune reconstruction efficiency, poor tumor immune microenvironment simulation degree, low clinical consistency and the like of an existing model are overcome, and the method can be used for tumor immune treatment drug screening, microenvironment mechanism research and personalized tumor treatment scheme verification.

The invention provides a construction method and application of a blood sinus networked liver organ. According to the method, pluripotent stem cells are co-differentiated into HAND1 < + > mesoderm and endoderm, a front intestine rear endoderm containing KDR < + > mesoderm progenitor cells is generated, a liver endoderm with CD34 < + > / CD31 < + > endothelial progenitor cells is generated, and the blood sinus networked liver organ is formed. The liver organ has a functional blood sinus endothelial network penetrating through the liver cluster, and has the characteristic functions of low-density lipoprotein uptake, human serum albumin removal, mediated secretion of blood coagulation factor 8 and the like. The method provides a new-generation highly bionic research tool for liver disease mechanism analysis, drug screening and regeneration repair.

Described herein are compositions and methods for modifying eukaryotic cells, for example, to express a transgene of interest and / or to produce an expanded population of cells ex vivo. Using the compositions and methods of the disclosure, a population of eukaryotic cells, such as a population of pluripotent cells (e.g., CD34+ hematopoietic stem or progenitor cells) may be transduced to express a gene of interest by contacting the cells with a viral vector, such as a lentiviral vector, and a diblock copolymer, such as a diblock copolymer composed of a hydrophilic region and a hydrophobic region. For example, the diblock copolymer may be composed of polyoxyethylene (PEO) subunits and polyoxypropylene (PRO) subunits. Additionally, the compositions and methods described herein can be used to promote the proliferation or survival of a population of pluripotent cells (e.g., CD34+ hematopoietic stem or progenitor cells) ex vivo, for example, by contacting the cells with a diblock copolymer.

The invention provides an RNP compound, an electrotransfection composition and a pharmaceutical composition thereof, a CD34 + cell gene editing method using the composition, and a method for obtaining HBG gene target mutation in a CD34 + cell genome.

The invention provides compositions and methods for treating diseases associated with expression of CD19, e.g., by administering a recombinant T cell comprising the CD19 CAR as described herein, in combination with one or more B-cell inhibitors, e.g., inhibitors of one or more of CD10, CD20, CD22, CD34, CD123, FLT-3, ROR1, CD79b, CD179b, or CD79a. The disclosure additionally features novel antigen binding domains and CAR molecules directed to CD20 and CD22, and uses, e.g., as monotherapies or in combination therapies. The invention also provides kits and compositions described herein.

A reagent kit, staining method, and application for combining dual immunohistochemistry with elastin fiber staining. The reagent kit includes a combination of primary antibodies, consisting of anti-CK7 and anti-CD34 monoclonal antibodies from different species, such as rabbit anti-human CK7 monoclonal antibody and mouse anti-human CD34 monoclonal antibody. By selecting CK7 and CD34 as the primary antibody combination, the growth pattern of tumor epithelium can be accurately indicated, and the proliferation and thickening of alveolar septal fibrous tissue caused by tumors can be clearly revealed, as opposed to structural changes of alveoli induced by other factors. Combined with independent Victoria blue staining, the elastin fibers of the walls of arteries and veins, as well as the pleura of the lungs, can be clearly visualized, aiding in the assessment of the integrity of elastin fibers on the lung surface and in determining the extent of tumor infiltration.

The invention relates to the technical field of cell differentiation, in particular to a method for directionally differentiating iPSC into iNK. Specifically, the method for preparing the CD34 + hematopoietic stem / progenitor cell and the iNK on the shaking table comprises the following steps: forming an embryoid (EB) by the iPSC on the shaking table, and then collecting a single cell supernatant to obtain the iNK. According to the method, high-proportion CD34 < + > hematopoietic stem / progenitor cells are obtained by adjusting the factor concentration and collecting the supernatant and are further induced and differentiated into the iNK, and more than 100 iNKs can be harvested before amplification of one iPSC according to the iNK harvested in batches.

The invention discloses a brain glioma animal model construction method based on an AAV mediated immune microenvironment. The method comprises the following steps: selecting an immunodeficient mouse at the right age; human CD34 + hematopoietic stem cells are injected for immune system reconstruction; human CD45 + immune cells in the peripheral blood PBMC are detected; the method comprises the following steps: selecting tumor tissues with high-expression brain glioma marker genes, carrying out rapid qPCR (quantitative polymerase chain reaction) or targeted RNA-seq immunohistochemical multiple verification, and screening consistent tumor tissues; the method comprises the following steps: physically dicing tumor tissues, treating the tumor tissues with pretreatment liquid, digesting the tumor tissues into single-cell suspension, and inoculating the single-cell suspension into a mouse cranium; an AAV vector carrying a specific immune regulation gene is injected into the tail vein of the mouse, and the brain glioma immune microenvironment is regulated; when it is observed that the tumor tissue grows stably and has immune cell infiltration characteristics, construction of the brain glioma animal model is completed. Compared with an existing model, the brain glioma model which is more stable, more scientific and closer to the human immune background is provided, and the brain glioma model is suitable for related immunotherapy drug screening and mechanism research.

Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells and a culture medium composition thereof. Provided are a method for inducing differentiation of pluripotent stem cells into CD34+ hematopoietic stem / progenitor cells or NK cells and a culture medium composition thereof. The NK cells obtained using the provided culture method have high purity and good in vitro expansion effect; the yield of iNK cells is high, and a single iPSC can differentiate into approximately 2,000 NK cells; and the obtained iNK cells highly express CD16 (over 70%). As a result, the problem of low CD16 expression in iNK cells in the prior art, which requires genetic modification methods to solve, has been addressed.

The present invention relates to a method for treating aging, and in particular immunosenescence, restoring thymic function and regenerating the thymus in a subject in need thereof, comprising the administration of T cell progenitors comprising CD7+CD34- cells.

The invention relates to an immune Raman multicolor imaging kit for liver cancer prediction and evaluation, the immune Raman multicolor imaging kit comprises five Raman probes, and the Raman probes are composed of Raman balls and antibodies connected to the surfaces of the Raman balls; wherein the Raman sphere is formed by polymerizing a triple bond monomer and (i) acrylic acid or (ii) styrene and acrylic acid, the particle size is 10-100 nm, and the Raman shift is 1800-2800 cm <-1 >; antibodies of the five Raman probes are respectively selected from one of an AFP antibody, an HBX antibody, a GPC3 antibody, a GS antibody and a CD34 antibody. The immune Raman multicolor imaging reagent and kit provided by the invention can realize one-step marking and multiple color development of five liver cancer related protein markers in the same liver tissue slice. Compared with a multiple fluorescence immunohistochemical technology, the method has remarkable advantages in efficiency and precision. Experiments show that the detection result is consistent with the clinical diagnosis result, and the application potential is huge.

The integration procedure provides different elements that require in vitro amplification of HSPCs, preferably for about 48 hours, in the presence of hematopoietic cytokines as well as UM171 and stem cell regenerative factor 1, and the introduction of i) a system (e.g., a CRISPR / Cas9 system) that generates DNA double-strand breaks (DSBs) upstream of the transcription start site of the target gene, and ii) the introduction of a donor matrix comprising cDNA with homologous arms on both sides (left [LHA] and right [RHA]).

Provided herein, inter alia, are methods and compositions for treating diabetes mellitus comprising co-transplantation of an insulin-producing cell and a cell derived from a parathyroid gland (PTG), a CD34+ cell derived from a parathyroid gland, a CD34+ cell derived from a stem cell, or other progenitor cell-derived CD34+ cell.

The invention discloses a method for constructing astrocytes serving as a smoke disease model, and belongs to the technical field of crossing of stem cells and neuroscience. The method comprises the following steps: S1) reprogramming CD34 + cells in in-vitro PBMCs (peripheral blood mononuclear cells) of smoke disease patients carrying and not carrying RNF213p.R4810K mutation to obtain induced pluripotent stem cells; and S2) directionally inducing and differentiating the induced pluripotent stem cells into astrocytes through a neural progenitor cell way, wherein the obtained astrocytes are the astrocytes capable of being used as the smoke disease model. The astrocyte model prepared by the invention can be used for researching pathogenesis, nerve-blood vessel interaction process and blood-brain barrier (BBB) related functions of smoke diseases, and can be further applied to molecular typing of diseases and in-vitro function evaluation of candidate drugs.

Novel methods for producing self-renewing erythroblasts (SREs) from induced pluripotent stem cells (iPSCs) or CD34+ cells are provided. Also provided are genetically modified iPSCs or CD34+ cells having a constitutively active stem cell factor (SCF) receptor and a constitutively active Janus kinase 2 (JAK2) that can differentiate into enucleated red blood cells, self-renew, and expand in culture media for an extended period of time. In particular, the disclosed methods completely eliminate the need for SCF and Epo.

The present disclosure relates to methods, media and supplements for culturing target cells, such as hematopoietic stem and progenitor cells (HSPC). The methods, media, and supplements of the present disclosure may include one or more epigenetic modifiers within culture conditions to culture and / or expand target cells, such as HSPC or CD34 + cells, such as may be obtained, enriched, or isolated from infected / diseased or normal primary samples. Output populations of HSPCs amplified in the presence of one or more epigenetic modifiers may be suitable for use in downstream applications.

Multispecific molecules that include an anti-G6B antibody or binding fragment thereof, an anti- CD34 antibody or binding fragment thereof; and one, two or all of: an immune cell engager (e.g., selected from an NK cell engager, a T cell engager, a B cell engager, a dendritic cell engager, and a macrophage cell engager); a cytokine molecule; a modulator of a cytokine molecule; a stromal modifying moiety; or any combination thereof are disclosed. Additionally disclosed are nucleic acids encoding the same, and methods of treating conditions or diseases using the aforementioned molecules.

The present disclosure relates to methods for selecting CD34 + cells, cells selected by said methods and therapeutic uses of those cells. More specifically, the present disclosure relates to CD34 + cells selected based on VEGF expression levels and / or cell and / or exosome microRNA expression levels. Thus, the present disclosure provides in vitro methods of selecting CD34 + cells based on determining the amount of VEGF and / or identifying miRNAs expressed by a population of CD34 + cells, and selecting CD34 + cells accordingly.

The present invention discloses a method for recognizing CD34 immunohistochemical images of hepatocellular carcinoma and calculating microvessel density. This method addresses the problems of manual diagnostic methods, which are complex, time-consuming, and labor-intensive, and rely on extensive diagnostic experience. In order to more comprehensively assess the severity of hepatocellular carcinoma patients and guide subsequent treatment, the present invention employs a deep learning network to perform binary classification on images, which has strong generalization capabilities and high robustness. The generated probability heat map intuitively reflects the model recognition results. The calculation of microvessel density is highly interpretable, simple, and convenient, making it easy for doctors to operate and adjust.

The invention provides a liver cancer capillary density evaluation system based on a CD34 and GPC-3 combined marker. The liver cancer microvessel density evaluation system based on the CD34 and GPC-3 combined marker comprises a digital pathology scanning module, a tumor area recognition module, a microvessel feature extraction module, a spatial correlation analysis module, a molecular subtype integration module, a microvessel invasion evaluation module, a prognosis prediction module, a dynamic layering module, a report generation module and a treatment recommendation module. The liver cancer microvessel density evaluation system based on the CD34 and GPC-3 combined marker has the advantages that the limitation of single quantification of microvessels is broken through, multi-dimensional accurate analysis of the liver cancer microenvironment is achieved through blood vessel function typing and spatial topology modeling, and conversion from pathological diagnosis to individualized treatment is promoted.

The present application provides materials and methods for treating hemoglobinopathies. More specifically, the application provides methods for producing progenitor cells that are genetically modified via genome editing to increase the production of fetal hemoglobin (HbF), as well as modified progenitor cells (including, for example, CD34+ human hematopoietic stem cells) producing increased levels of HbF, and methods of using such cells for treating hemoglobinopathies such as sickle cell anemia and β-thalassemia.

The invention provides a biomarker for detecting anti-VEGF treatment typing of diabetic macular edema and extracellular vesicle protein for judging the severity of diabetic retinopathy, and the protein with remarkable expression difference is screened out by analyzing the expression difference of the extracellular vesicle protein in aqueous humor of a patient. Wherein C5 and CD34 can be used as biomarkers of DME anti-VEGF treatment typing, C5 can be used for judging the severity of diabetic retinopathy (DR), important guidance is provided for DME or DR personalized treatment, and the C5 and CD34 have wide clinical application prospects.

The present invention provides methods for engrafting a bone marrow organoid with CD34+ hematopoietic stem and progenitor cells. Compositions comprising the factors needed for engraftment are also provided.

Compositions and methods are provided for modifying eukaryotic cells, e.g., to express a transgene of interest and / or to generate a population of expanded cells ex vivo.SOLUTION: A method of transducing a population of eukaryotic cells, such as a population of pluripotent cells, to express a gene of interest by contacting the cells with a viral vector, e.g., a lentiviral vector, and a diblock copolymer, e.g., a diblock copolymer comprised of a hydrophilic region and a hydrophobic region. For example, the diblock copolymer can be composed of polyoxyethylene (PEO) subunits and polyoxypropylene (PRO) subunits. In addition, the compositions and methods of the invention can be used to promote the ex vivo expansion or survival of a population of pluripotent cells (e.g., CD34 + hematopoietic stem or progenitor cells), for example, by contacting the cells with a diblock copolymer.SELECTED DRAWING: None

The invention provides a method of producing a population of CD34+ hematopoietic precursor cells. The CD34+ hematopoietic precursor cells are used in methods of producing natural killer (NK), methods of inducing NK cell differentiation from pluripotent stem cells (PSCs), and methods of generating terminally differentiated hematopoietic cells from PSCs. The differentiation of immune cells such as NK cells from PSCs includes the use of a hemogenic endothelium induction cocktail that includes a WNT signaling pathway activator, a bone morphogenetic protein and / or a vascular endothelial growth factor. Also provided is a method of producing hematopoietic stem cells from pluripotent stem cells.

According to the isolated culture identification and key marker detection method for the down producing goat hair follicle stem cells, trypsin and EDTA are used for jointly digesting skin tissue, and high-activity cells are efficiently obtained; a culture medium containing various growth factors is adopted for culture, and cell proliferation and stemness maintenance are promoted. And identifying and integrating morphological observation, growth curve determination and clone formation experiment to accurately judge the characteristics of the cells. Immunofluorescent staining and real-time fluorescent quantitative PCR are applied to key marker detection, and expression of markers such as CK15, CD34 and beta1 integrin is detected with high sensitivity. The method is high in separation efficiency, accurate in identification and sensitive in marker detection, can provide sufficient high-quality cells for the research on the hair follicle stem cells of the cashmere goat, assists in deeply understanding the biological characteristics and the regulation mechanism of the cells, and is of great significance for improving the yield and quality of cashmere and promoting the development of related biotechnologies.

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

The invention relates to a nucleic acid aptamer for targeting CD34 positive orbit fibroblasts, which is characterized in that the nucleotide sequence of the nucleic acid aptamer is as shown in SEQ ID NO: 10. The nucleic acid aptamer provided by the invention has the advantages of high affinity, strong specificity, small molecular weight, good stability, simple synthesis, low cost, easy modification, no immunogenicity and the like, and can meet the requirements of rapid detection of thyroid-related ophthalmopathy at present.

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