39 results about "Megakaryocytic cell" patented technology

The present invention provides methods and compositions for the production of hematopoietic progenitor cells or endothelial progenitor cells from human pluripotent stem cells using a defined cell culture medium without the need to utilize feeder cells or serum. In some embodiments, differentiation is accomplished using hypoxic atmospheric conditions. The defined medium of the present invention may contain growth factors and a matrix component. The hematopoietic progenitor cells may be further differentiated into cell lineages including red blood cells, macrophages, granulocytes, and megakaryocytes. The endothelial progenitor cells may be further differentiated into endothelial cells. Also disclosed are screening assays for identification of candidate substances that affect differentiation of pluripotent stem cells into progenitor cells.

It is an object of the present invention to provide a method for efficiently preparing blood cells, such as mature megakaryocytes and platelets, from iPS cells in an in vitro culture system.The present invention provides a sac-like structure enclosing hematopoietic progenitor cells, which is obtained by inoculating iPS cells onto feeder cells and then culturing the iPS cells under conditions suitable for inducing the differentiation of hematopoietic progenitor cells. Moreover, the present invention also provides a method for producing various types of blood cells, which comprises culturing hematopoietic progenitor cells enclosed in the sac-like structure under conditions suitable for inducing the differentiation of blood cells. Furthermore, the present invention also provides a method for producing various types of blood cells, particularly megakaryocytes and platelets, without involving the sac-like structure.

The invention relates to methods of obtaining compositions for generating multipotent hematopoietic stem progenitor cells comprising expansion of hematopoietic stem cells in the presence of HDACI and IDM. Methods of obtaining compositions enriched in hematopoietic megakaryocyte progenitor cells are also provided. Compositions enriched for stem cells and populations of cells obtained therefrom are also provided by the invention.

The invention provides an application of single cell sequencing in immune cell analysis. The immune cells comprise any one or a combination of at least two of T cells, mononuclear cells, natural killer cells, B cells, macrophages, granulocytes, mast cells, megakaryocytes or dendritic cells. Wherein the immune cell analysis comprises analysis of any one or a combination of at least two of types, states or proportions of immune cells and / or immune cell subtypes. In the present invention, single cell sequencing is applied to the immune cell analysis, a method comprises the following steps: firstly, obtaining a sequencing result of a sample through single cell sequencing, then obtaining a distribution state, subtype composition and a cell track of immune cells by adopting a biological information analysis method, annotating the cell subtypes, and realizing analysis on the types, states and proportions of the immune cells and / or the immune cell subtypes by comparing different results.

The present invention discloses the signaling pathway involved in erythroid repression by iron deficiency. Further disclosed is a non-toxic small-molecule compound which potently reverses the erythroid repression caused by iron deficiency. The present invention further encompasses novel compounds for inhibition of red cell production, useful, for example, in the treatment of polycythemia vera, a malignancy causing uncontrolled red cell production. These inhibitory compounds also promote megakary-ocytic lineage commitment and may therefore be useful for augmentation of platelet production. The present invention further discloses isocitrate reversal of iron deprivation.

A column-based device and method for retrieving cells of interest were enclosed. The said device comprises a column comprising (i) an inner wall defining an inner chamber with inlet and outlet openings, (ii) a perforated plug disposed adjacent to the outlet opening, (iii) a sleeve insert with a channel and disposed within the chamber and adjacent to the perforated plug, and (iv) a filtering means housed within sleeve insert sandwiched between two sealing means. In particular, Tumor-derived endothelial cell clusters (TECCs) as characterized multiple nuclei, expression of endothelial markers (PECAM1, VWF and CDH5), and non-expression of leukocyte, megakaryocyte and platelets markers, may be retrieved using the disclosed device. Also encompassed are methods, reagents and kits for the diagnosis and prognosis of cancers by detecting for the presence of TECCs isolated from blood samples using the claimed device.

The present invention relates to mainly one system of efficient culturing and directive inducing stem cell differentiation for culturing and proliferating megakaryocyte. The system has umbilical cord blood and marrow stem cell as seed cell, and the serum-free culture medium comprising TPO, IL-11 and heparin as the main component for megakaryocyte proliferation. The megakaryocyte preparation prepared based on the present invention contains great amount of macronucleus ancestral cells and mature megakaryocyte as well as CD34+ cells, and has the functions of treating thrombocytopenia and improving the blood-forming function after stem cell transplantation. The present invention provides novel efficient cell treating preparation for various thrombocytopenia and hemocyitopenia.

The invention discloses a therapeutic dendritic cell cancer vaccine and application thereof, and belongs to the technical field of biology. A shuttle plasmid pGBC-IRES-WSL and a recombinant adenoviruscapable of simultaneously expressing a tumor-associated antigen, a granulocyte-megakaryocyte colony stimulating factor (GM-CSF), CD40L and 4-1BBL is constructed, and human peripheral blood-derived mononuclear cells are infected with the recombinant adenovirus to obtain the therapeutic dendritic cell cancer vaccine. The cancer vaccine can play an anti-tumor role by inducing an organism to generateantigen-specific T lymphocytes so that the cancer vaccine is used for treating various blood system tumors and solid tumors.

A method for counting megakaryocytes in a specimen is described. In the method, first, erythrocytes in the specimen are lysed and nucleic acid in the megakaryocytes is stained with a fluorescent dye, and thereby, a measurement sample is prepared. Next, the cells in the measurement sample are irradiated with excited light so that the forward scattered light, the side scattered light and the fluorescence, which are emitted from the cells, are detected. Megakaryocytes are identified on the basis of the detected forward scattered light, the fluorescence and the side scattered light. Then, the identified megakaryocytes are counted.

The invention discloses a product for treating hemophilia A and an application thereof. The product comprises an expression cassette of a BDD FVIII gene, a recombinant vector containing the expressioncassette, a recombinant virus, especially a recombinant lentivirus, a recombinant bacterium or a recombinant cell. A promoter of the BDD FVIII gene in the expression cassette comprises a PF4 promoter. According to the lentiviral vector containing the PF4 promoter and the BDD FVIII gene expression cassette, the BDD FVIII gene can be specifically expressed in platelets formed by cytoplasm sheddingof megakaryocytes derived from CD34+ cells through the PF4 promoter, so that the blood coagulation function is achieved, and the lentiviral vector can be applied to gene therapy of hemophilia A of allages.

The present invention addresses the problem of providing a novel storage liquid for storing mammalian cells over a long period of time in a non-frozen state. Ascorbic acid and / or niacin are added to an isotonic liquid to give a storage liquid. Then the storage liquid is mixed with platelets and stored under shaking. Thus, deterioration in the functions of the platelets and a decrease in the survival rate thereof can be prevented for at least 10 days. When the aforesaid storage liquid is mixed with mesenchymal stem cells, megakaryocytes or T cells and stored in a non-frozen state, deterioration in the functions of these cells and a decrease in the survival rate thereof can be prevented for at least from several days to several ten days.

The invention relates to the technical field of biological cell preparation, in particular to a preparation method and application of an umbilical cord mesenchymal stem cell nutrient solution. The invention aims to provide a cell nutrient solution which can well repair skin cells and has a good beautifying effect and an application method of the cell nutrient solution. According to the technical scheme, the method comprises the following steps: separating mesenchymal stem cells, culturing the stem cells, and preparing a cell nutrient solution; the mesenchymal stem cell nutrient solution obtained by the invention comprises high-concentration human umbilical cord blood mesenchymal stem cells; a supernatant of the kit contains vascular endothelial cell growth factors, basic fibroblast growth factors, epidermal growth factors, IL-6 and IL-7, megakaryocyte colony stimulating factors, tumor necrosis factors, interferon and other factors, deoxyribonucleic acid and ribonucleic acid, monocyte chemotactic proteins, intercellular adhesion molecules, active polypeptides, immune proteins IgG, IgA, IgM, IgD, IgE and the like. The repair of tissue defects can be well promoted.

The present invention discloses the signaling pathway involved erythroid repression by iron deficiency. Further disclosed is anon-toxic small-molecule compound which potently reverses the erythroid repression caused by iron deficiency. The present invention further encompasses novel compounds for inhibition of red cell production, useful, for example, in the treatment of polycythemia vera, a malignancy causing uncontrolled red cell production. These inhibitory compounds also promote megakaryocytic lineage commitment and may therefore be useful for augmentation of platelet production. The present invention further discloses isocitrate reversal of iron deprivation.

The invention concerns a method for increasing megakaryocyte and megakaryocyte progenitor numbers in vitro or in vivo by suppressing SH2-containing inositol-5-phosphatase (SHIP) function in megakaryocytes or megakaryocyte progenitors expressing the SHIP gene. SHIP function can be suppressed by administering an interfering RNA, or other SHIP inhibitor, to the megakaryocytes or megakaryocyte progenitors in vitro or in vivo.

The present invention relates to novel cytoplasmic tyrosine kinases isolated from megakaryocytes (megakaryocyte kinases or MKKs) which are involved in cellular signal transduction pathways and to the use of these novel proteins in the diagnosis and treatment of disease. The present invention further relates to specific megakaryocyte kinases, designated MKK1, MKK2 and MKK3, and their use as diagnostic and therapeutic agents.

The invention relates to a tissue specific promoter and application thereof, wherein the nucleic acid sequence of the tissue specific promoter comprises more than 80% of sequences as shown in SEQ ID NO: 1, SEQ ID NO: 2, SEQ ID NO: 3 or SEQ ID NO: 4. The tissue specific promoter provided by the invention can start specific expression of coding genes in endothelial cells or megakaryocyte-platelet cells, can be applied to gene therapy requiring specific expression genes in endothelial cells or megakaryocyte-platelet cells, ensures the therapeutic effect, reduces the risk of immunological rejection, and saves the therapeutic cost.

An object of the present invention is to provide a separation substrate having a high megakaryocyte blocking rate and a high platelet permeation rate, and a cell separation filter and a method for producing a platelet which use the same. The separation substrate of the present invention is a separation substrate including a porous membrane for separating a platelet from a cell suspension containing a megakaryocyte and the platelet, in which an average pore diameter of the separation substrate is 2.0 μm to 12.0 μm, and the separation substrate is formed of at least one resin selected from the group consisting of a polysulfone resin and a polyvinylidene fluoride resin.

An object of the present invention is to provide a method for more efficiently differentiating CD34+ cells derived from umbilical cord blood into megakaryocytic lineage cells and for generating platelets. Coculture of CD34+ cells derived from umbilical cord blood with immortalized stromal cells in the presence of cytokines has enabled cell proliferation up to the 100,000-fold level, which has been impossible to achieve to date.

The invention discloses an application of AMF (amifostine) in treatment of tumors with high Cyclin D1 expression. According to research, AMF inhibits tumor cell growth by inhibiting cancer gene CyclinD1 expression in a targeted manner and resists the tumors by inhibiting the CyclinD1 expression in a targeted manner, and the functions are successfully verified through megakaryocyte leukemia cell lines Dami; and AMF successfully cures mantle cell lymphoma with high Cyclin D1 expression and high leukocyte expression as characteristics in clinical practice.

The invention relates to three megakaryocyte subgroups with different functional heterogeneity possibly existing in mouse megakaryocyte (MK), and specific molecular markers of the subgroups are respectively identified, including an MK molecular marker MYLK4 as a microenvironment supporting cell of HSC; the MK molecular marker for performing the immune regulation function is LSP1, CD53 or CD52; and the MK molecular marker with the plate production function is ARNTL. The molecular markers can be used for separating or marking MKs with different functions so as to promote the MKs to perform different functions, including HSC resting supporting, immunoregulation, plate production and the like.

The invention relates to construction of an SV40LT gene-mediated megakaryocyte model and a cell bank of megakaryocyte, belonging to the field of medical research. The construction is mainly characterized in that an SV40LTag-pcDNA3.1 (-) recombinant is constructed from T4DNA ligase, BamHI, pcDNA3.1 (-) DNA and SV40LTag DNA by virtue of conventional means; the recombinant is purified by virtue of competent escherichia coli; the recombinant is introduced into megakaryocyte cultured in vitro through a lipofection transfection method, so that the recombinant is integrated with the DNA of the cell; a cell screened by virtue of G418 and containing a positive recombinant is subjected to subculture and enlarged cultivation; a cell which is consistent with the biological properties of an immortalized cell in cellular morphology, cell growth curve, karyotype, nude mice carcinogenicity test, detection of SV40 large T gene in transfection cell DNA, determination of an mRNA expression product and a DNA sequence determination result is screened out as a megakaryocyte model, and is cryopreserved in liquid nitrogen, so as to construct an SV40LT gene-mediated megakaryocyte model and the cell bank of megakaryocyte; therefore, a method for long-term in vitro subculture of the megakaryocyte is established, and the method is applicable to the in vitro research of development, function and pathogenic mechanism of the megakaryocyte.

A process for separating megakarylcyte stimulator (MES) from human plasma and purifying it includes dissolving the deposite of human plasma, which is generated by extracting albumin and globulin centrifugal treating, ultrafiltrating, ion-exchange cellulose chromatography, gel filter and DEAE gel chromatography. The said MES has 40-42 Kda or 18-20 kda of molecular weight, and can promote the generation of thrombocytes and the multiplication of naval blood's stem cells, so it can be used for treating thrombocytopenia and stem cell transplant.

The present invention provides bio-nanoparticles (BioNPs) for delivering an active agent into hematopoietic stem & progenitor cells (HSPCs). Each BioNP comprises a core and a biological membrane covering the core, which comprises the active agent and a polymer. The biological membrane comprises a phospholipid bilayer and one or more surface proteins of a megakaryocyte (Mk). The active agent remains active after being delivered into the HSPC. Also provided are methods for preparing the BioNPs and uses of the BioNPs for targeted delivery of an active agent into HSPCs and / or treating or preventing a disease or condition in a subject in need thereof.

The present invention addresses the problem of providing: a separation substrate which has a high inhibition rate of megakaryocytes and a high permeability of platelets; a cell separation filter usingthe same; and a platelet producing method. The separation substrate of the present invention is composed of a non-woven fabric for separating platelets from cell suspension including megakaryocytes and platelets, wherein the average pore diameter of the separation substrate is 2.0-15.0 [mu]m and the thickness of the separation substrate is 10-500 [mu]m.

The invention discloses an application of histone methyltransferase inhibitor to preparation of a product for promoting megakaryocyte proliferation or blood platelet formation. The histone methyltransferase inhibitor is a histone methyltransferase G9a inhibitor. The histone methyltransferase G9a inhibitor stated in the invention can be used for promoting single CD34+hematopoietic stem and progenitor cells to differentiate and form 400 CD41a+CD42b+platelet granules, and the platelet granules are increased by 10 times than the platelet granules of a control group; the thrombopoiesis efficiency is further higher than the general range in literature report obviously, and a foundation is laid for mass production of functional platelets for clinic treatment in the future.

It has been demonstrated that GABRR1 is expressed on a subset of hematopoietic stem cells (HSC) and megakaryotic progenitor cells (MkP). MkP differentiation and reduction of the number of platelets in blood can be inhibited by inhibiting the GABRR1. GABRR1 overexpression or agonist treatment can significantly promote the generation of MkP and the growth of megakaryocytes.

An object of the present invention is to provide a separation substrate having a high megakaryocyte blocking rate and a high platelet permeation rate, and a cell separation filter and a method for producing a platelet which use the same. The separation substrate of the present invention is a separation substrate including non-woven fabric for separating a platelet from a cell suspension containing a megakaryocyte and the platelet, in which an average pore diameter of the separation substrate is 2.0 μm to 15.0 μm, and a thickness of the separation substrate is 10 μm to 500 μm.