37 results about "Megakaryocytic cell" patented technology

The invention discloses an intelligent bone marrow megakaryocyte morphology detection method which comprises the following steps: S1, data preparation: collecting and preprocessing a digital large map of a bone marrow smear, labeling megakaryocytes, and establishing a labeled sample; s2, generating and sorting a sample: extracting a small graph sample by taking megakaryocyte as a center; s3, constructing a deep learning model: constructing and training a deep convolutional neural network, and performing model optimization and performance improvement by using the generated small image sample set; s4, large image detection and reasoning: calling the trained model for reasoning by adopting a sliding window mechanism, and fusing detection results of a plurality of small windows back to an original large image through confidence weighting and a non-maximum suppression strategy; and S5, target cell segmentation: carrying out target segmentation on the detected megakaryocyte, and introducing a pyramid structure for cells with different sizes to obtain an accurate segmentation mask of each target cell. According to the method, the megakaryocyte detection and segmentation precision is improved through large image labeling, small image training and a sliding window reasoning strategy.

The invention relates to the field of biological medicine, and provides an application of a dihydroorotate dehydrogenase inhibitor in preparation of megakaryocytes, and the dihydroorotate dehydrogenase inhibitor is a compound shown as a formula I, a formula II or a formula III, or a pharmaceutically acceptable salt of the compound. According to the technical scheme, the dihydroorotate dehydrogenase inhibitor can promote hematopoietic stem / progenitor cells to differentiate into megakaryocytes and platelets, the platelet generation amount is increased by about 2 times, and a promising strategy is provided for in-vivo and in-vitro megakaryocyte and platelet generation.

The invention provides an application of a histone deacetylase (HDAC) inhibitor and / or a JAK2 inhibitor in inducing hematopoietic stem progenitor cells, mononuclear cells or total nucleated cells to generate megakaryocytes and / or megakaryocyte progenitor cells. The invention also provides methods of inducing hematopoietic stem progenitor cells, mononuclear cells or total nucleated cells to produce megakaryocytes and / or megakaryocyte progenitor cells comprising exposing the cells to an HDAC inhibitor, and / or exposing the cells to a JAK2 inhibitor.

The present disclosure provides, in various aspects and embodiments, methods for generating hematopoietic lineages for cell therapy, including erythroid progenitor cells, progenitor cell erythroblasts, granulocyte-macrophage progenitor cells (GMP), and megakaryocyte erythroid progenitor cells (MEPs) and erythroid cells. In various embodiments, the invention provides efficient in vitro methods for developing such hematopoietic lineages, including but not limited to progenitor erythroblasts and erythroblasts lineages, from human induced pluripotent stem cells (iPSCs). The cells produced in various embodiments according to the present disclosure are functional and / or more closely similar to corresponding lineages isolated from peripheral blood or bone marrow. The invention also provides isolated cells and cell compositions produced by the methods disclosed herein, as well as methods for cell therapy.

The invention discloses an application of a natural product WithangatinA in preparation of erythroleukemia induced differentiation treatment drugs, the natural product WithangatinA promotes HEL and K562 cells to be transformed from malignant proliferative cells to normal megakaryocytes by inducing the cell particle size to become larger and significantly increasing multinuclear megakaryocytes and promoting expression of surface labeled molecules differentiated from megakaryocytes at the same time, and the erythroleukemia induced differentiation treatment drugs are obtained. The effect of resisting erythroleukemia is achieved. After the natural product Withangulin A is adopted for treatment, the size and the weight of the spleen of a leukemia mouse can be remarkably reduced, the hematocrit and the platelet number of the mouse are remarkably increased, in addition, treatment of the natural product Withangulin A has no obvious influence on the weight of the mouse, and the safety is high. In-vivo research shows that the medicine can induce megakaryocyte differentiation of erythroleukemia in a mouse body, the effect of resisting erythroleukemia is achieved, and a research basis is provided for development of differentiation treatment medicines of erythroleukemia.

The present disclosure relates to the field of biological medicine, and provides a use of a dihydroorotate dehydrogenase inhibitor in the preparation of megakaryocytes, wherein the dihydroorotate dehydrogenase inhibitor is a compound represented by formula I, formula II or formula III, or a pharmaceutically acceptable salt of the compound. The dihydroorotate dehydrogenase inhibitor in the technical solution of the present disclosure can promote the differentiation of hematopoietic stem / progenitor cells into megakaryocytes and platelets, and increase the platelet production by about 2 times, thereby providing a promising strategy for the production of megakaryocytes and platelets in vivo and in vitro.

The invention belongs to the technical field of cell biology, and particularly relates to a cell culture method based on self-assembly microgel, a cell product and application. The cell culture method based on the self-assembled microgel comprises the following steps: culturing cells by adopting a cell culture medium, and adding the self-assembled microgel into a culture system. The cell culture method has the beneficial effects that the controllable and reversible self-assembly microgel scaffold is constructed by taking microgel with self-assembly capability as a basic construction unit, and three-dimensional wrapping, static culture and dynamic culture aiming at suspension cell types are realized in vitro; the cells obtained by the culture method can be transplanted in vivo, can be used for a clinical treatment scheme for assisting transplantation of cells in marrow cavities such as allogeneic hematopoietic stem cells and megakaryocytes, and can be used for optimizing cell therapy of hematological malignant tumors, thrombocytopenia and other hematological system malignant diseases.

The invention provides a method for producing platelets through human iPSCs (induced pluripotent stem cells) in-vitro differentiation, aiming at the existing in-vitro induction method, a systematically optimized in-vitro culture scheme is adopted, and the method mainly comprises the following four aspects: (1) optimizing forming conditions of embryoid bodies (EB); (2) optimizing the composition of a basic cell culture medium; (3) establishing an induction scheme for replacing cell factors with small molecular compounds; and (4) adding a small-molecule inhibitor to promote maturation and differentiation of megakaryocytes. The efficiency is improved by a technology for promoting MKs maturation by adjusting the amount of initial EB cells and adding small molecules, the cost is reduced by a method for optimizing culture medium components and replacing cell factors with small molecules, and the differentiation time of megakaryocytes is shortened to 14 days; the differentiation efficiency of the MKs is remarkably improved by about 2.5 times (about 1.428 megakaryocytes are obtained by differentiating each iPSC, and about 0.57 megakaryocytes are obtained by differentiating each iPSC according to a basic scheme), and the final differentiation maturity is remarkably improved. Meanwhile, after the cell culture medium is optimized, the production cost is obviously reduced by 70.9%.

The application belongs to the technical field of biology, and discloses a kit for inducing stem cells to differentiate into megakaryocytes and / or platelets and application thereof. The kit can induce stem cells to differentiate into megakaryocytes and / or platelets without introducing exogenous cells and exogenous genes. The kit can highly simulate the generation process of megakaryocytes and platelets in vivo, and can induce stem cells into mesendoderm cells, hematopoietic endothelial cells and megakaryocyte-erythroid progenitor cells in sequence. The megakaryocyte-erythroid progenitor cells further develop into megakaryocytes with a multiploid characteristic, and finally produce stem cell-derived platelets with similar functions to natural platelets. The stem cells have the potential for unlimited proliferation, and can be massively expanded in vitro, which provides an unlimited cell source for the in vitro preparation of platelets.

GABRR1 is shown to be expressed on subsets of hematopoietic stem cells (HSCs) and megakaryocyte progenitors (MkPs). Inhibition of GABRR1 inhibits MkP differentiation and reduction of platelet numbers in blood. Overexpression of GABRR1 or treatment with agonists significantly promotes MkP generation and growth of megakaryocytes.

The present application provides the use of leukotrienes in inducing stem cells to differentiate into hematopoietic stem / progenitor cells and the application thereof. According to specific embodiments of the present application, the addition of appropriate amounts of leukotrienes during the process of inducing stem cells to differentiate into hematopoietic stem / progenitor cells can significantly improve the differentiation efficiency of the stem cells into hematopoietic stem / progenitor cells, and significantly increase the number of erythroid cells, megakaryocytic cells, granulocytic cells and mononuclear cells obtained after the differentiation of the stem cells through hematopoietic stem / progenitor cells.

Provided are a tissue-specific promoter and use thereof. The tissue-specific promoter has a nucleic acid sequence comprising more than 80% of the sequence as shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. The tissue-specific promoter can promote the specific expression of a coding gene in endothelial cells (ECs) or megakaryocyte-platelet cells and can be applied to gene therapy in which a gene is required to be specifically expressed in ECs or megakaryocyte-platelet cells, ensuring a therapeutic effect, reducing a risk of immune rejection and saving a therapeutic cost.

The invention belongs to the field of platelet differentiation, and particularly relates to application of an adenosine A2a receptor stimulant in induced differentiation of platelets. The invention provides an application of an adenosine A2a receptor agonist in inducing differentiation of hematopoietic stem progenitor cells and / or megakaryocyte progenitor cells to generate platelets. The adenosine A2a receptor agonist contains Redelbrusone; in addition, the invention further provides a culture medium containing the adenosine A2a receptor stimulant and application of the culture medium in preparation of platelets through in-vitro differentiation, and provides an in-vitro differentiation method of the platelets, and the in-vitro differentiation method of the platelets improves the yield of the platelets by nearly one time.

Cell-derived compositions having physiological activity are provided. The compositions of the invention comprise a megakaryocyte or a treatment of a culture thereof.

This application discloses a method, system, device, storage medium, and product for detecting megakaryocytes, comprising: acquiring pathological slide images and preprocessing the pathological slides, wherein the preprocessing further includes generating small-sized pathological slides based on the pathological slides; inputting the preprocessed pathological slides into a pre-trained segmentation model to output megakaryocyte information and location information; stitching the small pathological slides to generate a high-resolution pathological slide image; inputting the high-resolution pathological slide image into a classification model to output megakaryocyte classification results; acquiring megakaryocyte clustering information based on the high-resolution pathological slide image, and generating medical information based on the megakaryocyte information and location information, the megakaryocyte classification results, and the megakaryocyte clustering information. This method solves the problems of low accuracy and long reading time in manual review of megakaryocyte slides.

Methods for producing megakaryoctyres 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 invention discloses a sound tweezer method and circuit control system for megakaryocyte sorting, and the method comprises the following steps: step 1, providing a cell sorting chip which comprises a PDMS flow channel, a two-way sorting flow channel and a three-way sorting flow channel, and the PDMS flow channel is internally provided with a cylindrical array; step 2, injecting a sample solution containing cells or particles with different diameters into the PDMS flow channel; 3, after the cells or the particles are subjected to cylindrical array sorting in the PDMS flow channel, the cells or the particles with the diameters smaller than the critical diameter are guided into a two-separation flow channel from the PDMS flow channel, and the cells or the particles with the diameters larger than the critical diameter are guided into a three-separation flow channel from the PDMS flow channel; and 4, sound fields are applied to the two-way sorting flow channel and the three-way sorting flow channel, and the cells or particles with different diameters enter different outlets respectively by adjusting sound field parameters, so that multi-scale sorting is realized. The multi-scale megakaryocyte sorting device can realize multi-scale megakaryocyte sorting, and has the advantages of high efficiency, stability and convenience in operation.

Methods, compositions, and systems for treating subject(s) in need of plasma Factor VIII, particularly a subject having preexisting anti-FVIII inhibitory antibodies, are provided. The methods involve administering to the subject a therapeutically effective amount of an inflammation suppressor, a therapeutically effective amount of a CD8+ T cell depleting agent, and a therapeutically effective amount of a composition comprising a lentiviral vector (LV) comprising an optimized FVIII expression cassette expressibly linked to a megakaryocyte-specific promoter. Such methods, compositions, and systems are useful to treat subjects with blood clotting disorder(s), such as hemophilia A.

Methods for producing megakaryoctyres and platelets derived from inducible pluripotent stem cells and comprising a therapeutic agent are provided. 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 invention provides an anti-human CD42b antibody and application thereof. The antibody comprises a heavy chain variable region and a light chain variable region, the CDR1, the CDR2 and the CDR3 of the heavy chain variable region are amino acid sequences as shown in SEQ ID NO. 1, SEQ ID NO. 2 and SEQ ID NO. 3 respectively; the CDR1, the CDR2 and the CDR3 of the light chain variable region are amino acid sequences as shown in SEQ ID NO. 4, SEQ ID NO. 5 and SEQ ID NO. 6 respectively. The anti-human CD42b antibody provided by the invention provides an anti-human CD42b monoclonal antibody light chain variable region gene and a heavy chain variable region gene, and a hybridoma cell strain capable of generating a CD42b specific antibody, and can be used for detection of human and megakaryocyte / platelets.

The present application relates to a method for improving the quality of platelets produced in vitro. Specifically, the present inventors provide a platelet culture medium for in vitro culture, which can significantly improve the yield and quality of platelets cultured in vitro by optimizing the ingredients and adding new ingredients. Specifically, the viability of platelets obtained by culturing megakaryocytes in the culture medium of the present application and the proportion of CD42b+ cells are significantly increased, and the number of platelets produced by a single megakaryocyte is also significantly increased. The present application improves the problems of imperfect platelet function and unstable quality in existing in vitro platelet production technology, provides a stable platelet source for clinical transfusion, and has a wide application prospect.

Provided is a use of a histone deacetylase (HDAC) inhibitor and / or JAK2 inhibitor in inducing hematopoietic stem progenitor cells, mononuclear cells or total nucleated cells to generate megakaryocytes and / or megakaryocyte progenitor cells. Also provided is a method for inducing hematopoietic stem progenitor cells, mononuclear cells or total nucleated cells to generate megakaryocytes and / or megakaryocyte progenitor cells, comprising exposing the cells to an HDAC inhibitor and / or exposing the cells to a JAK2 inhibitor.

Methods for producing megakaryocytes and platelets derived from inducible pluripotent stem cells and comprising a therapeutic agent are provided. 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.

This disclosure provides reagents and methods for producing NK progenitor cells, NK cells, megakaryocytes and CAR derivatives thereof from hematopoietic stem cells. Pharmaceutical compositions comprising NK progenitor cells produced by the reagents and methods of the invention and immunotherapeutic methods using these pharmaceutical compositions are also provided.

The invention relates to a method for improving the quality of platelets produced in vitro. Specifically, the inventor provides the in-vitro culture platelet culture medium, and the yield and the quality of the in-vitro culture platelet can be remarkably improved by optimizing components and adding novel components. Specifically, when megakaryocytes are cultured by adopting the culture medium disclosed by the invention, the motility rate of obtained platelets and the proportion of CD42b + cells are remarkably increased, and the number of platelets generated by a single megakaryocyte is also remarkably increased. According to the invention, the problems of incomplete platelet function, unstable quality and the like in the existing in-vitro platelet production technology are solved, a stable platelet source is provided for clinical infusion, and the application prospect is wide.

The present invention provides a method for producing human immortalized megakaryocytes and / or human immortalized megakaryocyte progenitor cells and / or human megakaryocyte cell lines (102) from adult CD34 + HSPC (100) capable of maturation into functional megakaryocytes (103) producing functional platelets (104), said method comprising at least the steps of: producing functional platelets (104) by transduction, in particular lentiviral (LV) transduction; an expression cassette comprising at least one gene is introduced into the CD34 + HSPC (100) to effectuate forced expression of the at least one gene wherein the transduced CD34 + HSPC (100) is configured to express at least one gene from the HOXL subclass of the ANTP homologous cassette family and / or the TALE homologous cassette family upon activator-induced expression, and constitutively expressing at least one anti-apoptotic gene from the BCL2 family wherein the transduced CD34 + HSPC (100) is immortalized by expression of the at least one gene from the ANTP homologous cassette family and / or the HOXL subclass of the TALE homologous cassette family, preferably wherein the at least one gene from the HOXL subclass comprises a combination of one of the HOXB8, HOXA7, HOXA10 or HOXB4 genes and at least one of the MEIS1, MEIS2, MEIS3, PBX1, PBX2, PBX3 and / or PBX4 genes, and the at least one anti-apoptosis gene from the BCL2 family comprises BCL2L1 or BCL2L2. The invention further provides a method of producing functional platelets by using the immortalized megakaryocytes and / or immortalized megakaryocyte progenitor cells and / or immortalized megakaryocyte cell lines obtained by the above method. Further, the present invention provides a pharmaceutical composition comprising functional platelets obtained by the above method and its therapeutic use.

To provide compositions and methods for producing megakaryocytes.SOLUTION: There is provided a method for producing megakaryocyte precursor cells (preMK) and megakaryocytes (MK) from stem cells. The present disclosure further provides compositions comprising preMK and MK and lysates thereof, and also provides methods of using preMK, MK, lysates thereof, and compositions thereof. In one aspect, provided is a method for megakaryocyte production, the method comprising: expanding pluripotent stem cells under low adherent or non-adherent conditions and under agitation, the expanded pluripotent stem cells forming self-aggregating spheroids; differentiating the pluripotent cells in a first culture medium into hemogenic endothelial cells; and differentiating the hemogenic endothelial cells in a second culture medium into megakaryocytic progenitors.SELECTED DRAWING: None

The invention provides a combination capable of efficiently inducing differentiation of hematopoietic stem / progenitor cells to megakaryocyte lineage cells. The combination comprises three components: a first component, a second component and a third component, wherein the first component is an HSE7 gene or protein, mRNA of HSE7, an HSE7 expression cassette or an HSE7 accelerant; the second component is a histone deacetylase inhibitor; and the third component is a gamma-aminobutyric acid agonist. Also provided is a method for efficiently inducing differentiation of hematopoietic stem / progenitor cells into meganuclear lineage cells, comprising overexpressing HSE7, inhibiting HDAC expression and / or activity, and administering a GABA agonist.

Disclosed herein are compositions and methods related to megakaryocyte-derived extracellular vesicles derived from human pluripotent stem cells, where the megakaryocyte-derived extracellular vesicles may be utilized for treating Fanconi anemia (FA).