28 results about "Megakaryocyte" patented technology

The invention discloses a method for differentiating induced pluripotent stem cells into megakaryocytes and platelets, and relates to the technical field of induced pluripotent stem cell directional differentiation, and the method comprises the following steps: inoculating an iPSC single cell into a culture vessel, and carrying out adherent culture through a culture solution; after the cell adheres to the wall and before cloning is not formed, the liquid is changed into a first induction culture liquid, and the cell is induced and differentiated into a mesoderm cell; changing the solution into a second induction culture solution to induce and differentiate the mesoderm cells into hematopoietic endothelial cells; collecting the suspension cells, culturing the suspension cells in an amplification culture solution, and enriching megakaryocytes; inducing megakaryocyte polyploidy through a third induction culture solution; and culturing the polyploidy megakaryocyte through a fourth induction culture solution to promote platelet release. The method for inducing the pluripotent stem cells to differentiate into megakaryocytes and platelets is convenient and rapid, the megakaryocytes and platelets can be obtained through iPSC in a short time, and feeder cells, serum and additional gene editing are not needed; meanwhile, the yield is high.

The invention relates to the technical field of biology, in particular to a method for producing a platelet quality control product by using iPS cells and the platelet quality control product, and the method comprises the following steps: controlling overexpression of a specific gene in the iPS cells by using a gene editing technology to obtain an iPS cell line containing the specific iPS cells; a differentiation procedure is started by adding a culture medium containing a tetracycline derivative, and meanwhile, a fibroblast growth factor 2 and a bone morphogenetic protein 4 are added to promote mesoderm differentiation; growth factors and tetracycline derivatives are added to induce overexpression of specific gene segments and promote formation of specific megakaryocytes; adding an AhR antagonist and a ROCK inhibitor into the recovered specific megakaryocyte, and culturing to obtain platelets; and screening a platelet standard substance based on platelet functions and preparing the platelet standard substance into a quality control substance.

The invention provides a reagent composition for detecting minimal residual diseases of acute megakaryocytic leukemia and application of the reagent composition. The reagent composition comprises two groups of antibodies and is a flow cytometry detection composition for follow-up visit of minimal residual diseases after acute megakaryocytic leukemia treatment, a two-tube parallel scheme is used during application, and the first group of antibodies and the second group of antibodies are respectively used for different tube samples. The reagent composition can be applied to flow cytometry to detect minimal residual diseases after acute megakaryocytic leukemia treatment.

The present disclosure provides efficient in vitro methods for producing megakaryocytes and / or platelets from human induced pluripotent stem cells (iPSCs). In various embodiments, the cells and platelets produced in accordance with the present disclosure are functional and / or more closely similar to corresponding lineages isolated from peripheral blood, bone marrow, or other tissue. The present invention provides, in some aspects, isolated cells / platelets and compositions produced by the methods disclosed herein, as well as methods for use in therapy.

The invention discloses an application of a drug targeting a CCDC25 gene in preparation of a drug for preventing and / or treating tumor bone marrow metastasis related complications. The invention finds that CCDC25 is associated with the occurrence of tumor bone marrow metastasis related complications for the first time, and participates in the occurrence of tumor bone marrow metastasis related thrombocytopenia in the bone marrow metastasis process. After the CCDC25 is knocked out or subjected to targeted inhibition, differentiation of hematopoietic stem cells in bone marrow to megakaryocytes and the capability of mature megakaryocytes to generate platelets are effectively protected, which indicates that the knockout of the CCDC25 effectively improves thrombocytopenia caused by tumor bone marrow metastasis, which indicates that the CCDC25 can be used as a target for preventing and / or treating thrombocytopenia related to bone marrow metastasis. And the application value in research and development of medicines for treating thrombocytopenia related to bone marrow metastasis is great.

A method of selectively modulating gene expression of one or more target genes in megakaryocytes, circulating platelets, and / or platelets generated by the transfected megakaryocytes, in a subject in need thereof, wherein the method comprises administering to the subject a therapeutically effective amount of composition comprising naked RNA oligonucleotides.

The present application provides a kind of acute megakaryocyte leukemia micro residual disease detection reagent composition and its application.The reagent composition includes two groups of antibodies, and is used for the composition of flow cytometry detection for following-up micro residual disease after the treatment of acute megakaryocyte leukemia, and when used, 2 tubes are used in parallel, wherein the first group of antibodies and the second group of antibodies are used for different tube samples respectively.The reagent composition of the present application can be applied to flow cytometry detection of micro residual disease after the treatment of acute megakaryocyte leukemia.

Methods of production and apparatuses for production for stably producing healthy and functional platelets, and methods for determining operating conditions of an apparatus for producing platelets are provided. The present invention relates to methods of production comprising a step of culturing megakaryocytes in a platelet production medium C, wherein the step of culturing comprises a step of stirring a medium C in containers 1, 4 using stirring mechanisms 2, 5. In one method of production, the step of stirring comprises reciprocating stirring blades 21, 24 in the stirring mechanisms 2, 24 such that any of predetermined turbulent energy, shearing stresses, or Kolmogorov scale is produced, in the medium C. In another method of production, the step of stirring comprises pivoting a stirring blade 51 in the stirring mechanism 5 wherein a plurality of stationary members 53 is placed therearound in a bottom region 4e of the container 4 such that the step of stirring produces turbulence in the medium C. The present invention also relates to apparatuses for production for carrying out the method of production. The present invention also relates to a method for determining operating conditions of an apparatus for producing platelets.

The invention belongs to the technical field of biological medicines, and particularly relates to application of a compound Aglaroxin C in preparation of an erythroleukemia induced differentiation treatment medicine. The compound Aglaroxin C disclosed by the invention has the activity of resisting erythroleukemia cells HEL and K562, and the proliferation of the HEL and K562 cells is inhibited by inducing the HEL and K562 cells to generate G2 phase cycle arrest and cell apoptosis. According to the Aglaroxin C disclosed by the invention, the grain sizes of HEL and K562 cells are induced to become larger, multinuclear cells are remarkably increased, and meanwhile, the expression of a surface marker differentiated from a megakaryocyte line is promoted, so that the HEL and K562 cells are promoted to differentiate to the megakaryocyte line.

An embodiment of the present invention discloses a risk prediction system for an immune effector cell mediated neurotoxic syndrome (ICANS) and an application thereof, the system comprising: a processor and a memory coupled to the processor, the memory storing instructions, when the instructions are executed by the processor, the following steps are used: inputting a numerical value of an index into a prediction model to obtain an occurrence probability of the immune effector cell mediated neurotoxic syndrome; the prediction model comprises the following steps: by taking free DNA (cfDNA) traceability concentrations of oligodendrocytes A, B cells B and megakaryocytes C as prediction factors, establishing a risk prediction system formula of the immune effector cell mediated neurotoxic syndrome; the system can be used to assess the risk of immune effector cell mediated neurotoxic syndrome and possibly guide the selection of therapeutic regimens thereof.

The present application provides a method for inducing differentiation of embryonic stem cells into megakaryocytes. The provided method comprises the step of inducing differentiation of engineered embryonic stem cells into megakaryocytes; the engineered embryonic stem cells express a reduced amount of SHMT2 protein compared to unengineered embryonic stem cells. Megakaryocytes and platelets can be efficiently prepared via the method.

The present invention provides a human megakaryocyte-biased hematopoietic stem cell molecular marker and a detection kit thereof. The detection marker of the present invention fills the gap in the prior art for the identification of surface molecular markers of human megakaryocyte-biased hematopoietic stem cells, creatively discovers that megakaryocyte hematopoietic stem cells are enriched in CD34 + CD38 ‑ CD45RA ‑ CD90 + CD49f low Cell populations, using this marker to identify cell subpopulations or screen for the preparation of cell therapy preparations has broad application prospects and great improvements in scientific research and clinical practice.

Embodiments of the disclosure include systems, methods, and compositions for producing megakaryocytes and platelets for recipient individuals in need thereof. In certain embodiments the megakaryocytes and platelets may be produced following co-culture of MSCs and CD34+ cells in media comprising stem cell factor, thrombopoietin, IL-6, IL-3, FLT3-ligand, and an inhibitor of Rho associated coiled – coil containing protein kinase (ROCK).

The present invention relates to a method for regulating the degree of differentiation of cells, comprising a step for forcibly expressing an MYC family gene and a BMI1 gene in a cell selected from the group consisting of (i) mesenchymal stem cells, vascular endothelial cells, smooth muscle cells, neural crest cells, and lymphocytes having an arbitrary degree of differentiation, and (ii) megakaryocytes and megayoung granulocytes having a high degree of differentiation.

The purpose of the present invention is to provide a bone-forming composition comprising enlarged megakaryocytes and / or extracellular secretions derived from enlarged megakaryocytes.　The bone-forming composition according to the present disclosure comprises enlarged megakaryocytes and / or extracellular secretions derived from enlarged megakaryocytes.

The invention relates to a bone marrow smear test method for artificial intelligence assisted identification of small / micro megakaryocytes, which comprises the following steps: collecting bone marrow smears of MDS patients, performing full-piece scanning by using a 100X oil lens, positioning cells by using a target detection algorithm, performing artificial classification, amplifying the sample quantity of the small / micro megakaryocytes through treatment of brittleness, temprature and vibrice, and training the algorithm to obtain a cell classification algorithm; in the practical application, the whole scanning image of the bone marrow smear is obtained, the small / micro megakaryocyte is automatically identified by using the algorithm, and the manual auditing is performed on the basis of the algorithm result, so that the omission can be reduced, the integrity and accuracy of the inspection result are effectively improved, and the inspection efficiency of the bone marrow smear is improved.

This invention relates to the field of platelet production technology, and in particular to a method for producing platelets based on biomimetic microcarriers. The method overcomes the key bottlenecks of low in vitro platelet yield, incomplete function, and difficulty in large-scale production in the prior art. By constructing a biomimetic bone marrow-vascular wall microenvironment, it systematically solves the technical problem of low yield of megakaryocytes in in vitro culture systems, and provides a brand-new solution for the safe, efficient, and economical in vitro preparation of platelets.

The present invention provides a method for large-scale expansion of human hematopoietic stem cells based on bionic microcarriers. On the one hand, the large-scale culture can obtain a sufficient number of functional hematopoietic stem cells. On the other hand, the large-scale culture can maintain the stemness of hematopoietic stem cells for a long time and enhance their potential for differentiation into the megakaryocyte system, overcoming the defect that the absolute number of cells obtained by in vitro culture in the prior art is insufficient for treatment. At the same time, the method can directly culture human monocytes, directly collect and culture human monocytes, and avoid the unknown risks brought about by the process of sorting hematopoietic stem and progenitor cells. The method provided by the present invention takes into account the principles of efficiency and economy and has broad experimental and clinical application prospects. The present invention also provides the application of the large-scale culture method in the preparation of blood disease treatment preparations, and the application of the cell products obtained based on the large-scale culture method in the preparation of blood disease treatment agents.

The invention pertains to methods for the production of polyploid megakaryocytes using diploid megakaryocytes derived from induced pluripotent stem cells (iPSCs), the polyploid megakaryocytes so produced, and their use for the production of iPSC-derived platelet-like particles. The invention further pertains to the iPSCs-derived platelet-like particles produced using the polyploid megakaryocytes of the invention, and their use as a medicament.

We disclose a method, comprising extracting cells from a tissue of a donor, wherein the tissue is selected from the group consisting of blood and bone marrow; isolating CD14+ monocytes from the extracted cells; culturing the isolated CD14+ monocytes; differentiating the cultured CD14+ monocytes into megakaryocytes; and administering, to a patient suffering from a platelet deficiency, at least one cell type selected from the group consisting of the differentiated megakaryocytes and platelets produced by the differentiated megakaryocytes. We also disclose a kit comprising at least one material usable in a method; and instructions to perform a method.

The invention discloses a method for inducing hematopoietic stem / progenitor cells into myeloid progenitor cells and a culture medium used in the method. The method comprises a three-stage induced differentiation strategy of hematopoietic stem cells-pluripotent progenitor cells-myeloid progenitor cells, wherein a first-stage culture medium takes low-concentration cell factors and small molecule components as main components, and mainly promotes enrichment and amplification of the hematopoietic stem cells and enables the hematopoietic stem cells to smoothly transit to an in-vitro culture environment; the second-stage culture medium takes high-concentration cell factors and small molecules as main components and mainly promotes amplification of pluripotent progenitor cells and induced differentiation of common myeloid progenitor cells; in the third stage, the culture medium takes high-concentration cell factors, myeloid induction factors and small molecules as main materials to promote efficient amplification of three kinds of myeloid progenitor cells including common myeloid progenitor cells, granulocyte mononuclear myeloid progenitor cells and megakaryocyte erythroid myeloid progenitor cells, and finally a myeloid progenitor cell product is obtained. The myeloid progenitor cell amplification method provided by the invention greatly improves the induced amplification efficiency of the myeloid progenitor cells, and has very high clinical transformation application potential.

Methods for producing megakaryocytes and platelets derived from inducible pluripotent stem cells are provided. Such megakaryocytes or platelets can be genetically modified to comprise a nucleic acid molecule encoding a therapeutic agent. The present disclosure further provides methods and compositions for loading a platelet or a megakaryocyte with a therapeutic agent and for genetically modifying a platelet or a megakaryocyte to express an agent.

The present invention discloses the application of helenalin in the preparation of a medicament for alleviating and treating erythroid differentiation disorder of cells in a subject, belonging to the field of pharmaceutical preparations. The chemical structure of helenalin disclosed in the present invention is shown in Formula 1. It has been experimentally verified that helenalin can effectively correct the erythroid differentiation disorder caused by ribosomal protein gene mutations, specifically manifested as promoting the differentiation of hematopoietic stem cells with erythroid differentiation disorder into multipotent progenitor cells, promoting the differentiation of common myeloid progenitor cells with erythroid differentiation disorder into megakaryocyte-erythroid progenitor cells, alleviating problems such as the decrease in the proliferation ability of erythroid phenotype cells and the increase in apoptosis in erythroid differentiation disorder caused by ribosomal protein gene mutations, so that the erythroid phenotype cells are overall alleviated or restored. Therefore, the present invention can provide ideas for the treatment of diseases with erythroid differentiation disorder, such as DBA drugs.

The present invention provides methods for producing megakaryocytes and platelets derived from inducible pluripotent stem cells. Such megakaryocytes or platelets may be genetically modified to include a nucleic acid molecule encoding a therapeutic agent. The invention also provides methods and compositions for loading therapeutic agents to platelets or megakaryocytes and for genetically modifying platelets or megakaryocytes to express therapeutic agents.

The present invention provides a method of high-density cell culture comprising at least the following steps: S1: obtain hydrogel microcapsules through membrane emulsification; S2: encapsulate a selected number of cells per each obtained hydrogel microcapsule, and S3: culture the encapsulated cells in a cell culture medium. The invention further refers to Human Pluripotent Stem Cells (hPSCs) obtained through the above-described method, in particular for use in medicine, more in particular for use in a method of in vitro generation of mature megakaryocytes.

The invention discloses a method for detecting megakaryocytes in a bronchopulmonary dysplasia model, and relates to the field of biomedicine. Comprising the following steps: constructing a BPD mouse model, carrying out HE staining, flow cytometry and immunofluorescence detection on lung tissues, and analyzing the number, proportion, ploidy and distribution of megakaryocytes, the construction of the BPD mouse model comprises the following steps: randomly dividing C57BL / 6j strain newborn mice into an experimental group and a control group one day after the newborn mice are born, feeding the experimental group in a 85-90% high oxygen concentration environment for 14 days, and feeding the control group in a C57BL / 6j strain newborn mice in a C57BL / 6j strain newborn mice in a C57BL / 6j strain newborn mice. The control group is fed in the air, and the female rats are interchanged among the groups every 24 hours. By integrating model construction, HE staining, flow cytometry and immunofluorescence, multi-dimensional detection of megakaryocytes in the BPD model is achieved, the method is standardized and good in repeatability, reduction and ploidy change of megakaryocytes in lung tissue can be accurately reflected, and a reliable tool is provided for research of a BPD mechanism.

Provided is a method for selectively modulating gene expression of one or more target genes in megakaryocytes, circulating platelets, and / or platelets produced by transfected megakaryocytes in a subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of a composition comprising a naked RNA oligonucleotide.