36 results about "Erythroblast" patented technology

A method for discriminating and counting erythroblasts comprises the steps of: (i) staining leukocytes in a hematologic sample by adding a fluorescent labeled antibody capable of binding specifically with leukocytes to the hematologic sample; (ii) raising the permeability only of cell membranes of erythroblasts in the hematologic sample to a nucleotide fluorescent dye which does not permeate a cell membrane usually, the nucleotide fluorescent dye having a fluorescent spectrum capable of being distinguished from that of a fluorescent labeling compound of the fluorescent labeled antibody in step (i); (iii) staining nuclei of the erythroblasts in the hematologic sample with the nucleotide fluorescent dye; (iv) subjecting the hematologic sample to flowcytometry to detect at least two fluorescent signals from each cell; and (v) discriminating and counting the erythroblasts from difference in intensity between the at least two fluorescent signals.

Mithramycin derivatives and their pharmaceutically acceptable salts are disclosed. The mithramycin derivatives can be used in the treatment of Ewing sarcoma or other cancer or neuro-disease associated with an aberrant erythroblast transformation-specific transcription factor.

On one hand, the invention provides a compound with a structure of general formula I or a conjugate thereof, wherein each group is as defined in the description. On the other hand, the invention provides a composition comprising (i) the compound of formula I or the conjugate thereof, and (ii) at least one surfactant selected from cationic surfactants and nonionic surfactants. The invention also provides a preparation method of the composition and a kit containing the composition. The invention still provides a method for the simultaneous identification of nucleated erythrocyte, basophilic granulocyte and lymphocyte and the classification by using the composition disclosed in the invention.

A method for discriminating and counting erythroblasts comprises the steps of: (i) staining leukocytes in a hematologic sample by adding a fluorescent labeled antibody capable of binding specifically with leukocytes to the hematologic sample; (ii) raising the permeability only of cell membranes of erythroblasts in the hematologic sample to a nucleotide fluorescent dye which does not permeate a cell membrane usually, the nucleotide fluorescent dye having a fluorescent spectrum capable of being distinguished from that of a fluorescent labeling compound of the fluorescent labeled antibody in step (i); (iii) staining nuclei of the erythroblasts in the hematologic sample with the nucleotide fluorescent dye; (iv) subjecting the hematologic sample to flowcytometry to detect at least two fluorescent signals from each cell; and (v) discriminating and counting the erythroblasts from difference in intensity between the at least two fluorescent signals.

The invention relates to an application of miR-125b in red blood cell maturation. A method for preparing mature red blood cell comprises the following steps: making precursor cells of the red blood cell to express miR-125b, wherein the miR-125b has a nucleotide sequence shown in a SEQ IDNO:1; and culturing the precursor cells of the red blood cell of the expressed miR-125b to obtain the mature red blood cell; wherein, the precursor cells of the red blood cell can be selected from original erythroblast, polychromatic erythroblast, orthoneutrophil erythroblast and reticulocyte. The method can be used for effectively preparing the mature red blood cell.

We describe a new two-step culture method for mass production in vitro of erythroid cells from either CD34+ (105 cells / mL) or light-density (106 cells / mL) cells purified from the blood of normal donors and thalassemic patients. The method includes (i) culture of the cells in the presence of dexamethasone and estradiol (10−6 M each) and (ii) the growth factors SCF (50 ng / mL), IL-3 (1 ng / mL), and EPO (1 U / mL). In their proliferative phase, these cultures generated about 1-2×107 erythroblasts for each milliliter of blood collected from normal donors or thalassemic patients. They were composed mostly (90%) of CD45low / glycophorin (GPA)neg / CD71low cells at day 7, 50-60% of which became CD45neg / GPA+ / CD71high by days 15-20. However, when cells from days 7 to 12 of the proliferative phase were transferred in differentiation medium containing EPO and insulin, they progressed to mature erythroblasts (>90% benzidinepos and CD45neg / GPA+ / CD71medium) in 4 days. Because of the high number of erythroid cells that are generated from modest volumes of blood, this method will prove useful in donor-specific studies of erythroid differentiation.

Described herein is a method of differentiation of pluripotent stem cells into hematopoietic precursor cells, hematopoietic progenitor cells, erythroblast cells, and enucleated erythroblast cells, wherein the method is carried out under suspension agitation, and a GSK-3-inhibitor or a Wnt pathway activator is added during a stage of mesoderm induction. In the preferred embodiment, the pluripotentstem cells are expanded in agitated microcarrier culture and the GSK-3-inhibitior is CHIR99021. Also disclosed herein are cell culture media for use in the methods disclosed herein, as well as kits for performing the same.

The enumeration and analysis of residual white blood cells in a sample of leukocyte-reduced blood products is conducted by forming a suspension of the leukocyte-reduced blood products with a sufficient amount of a lysing reagent. The lysing reagent comprises a buffer with a low molar concentration, and a non-ionic surfactant. The suspension of leukocyte-reduced blood products and the lysing reagent is incubated for a sufficient time at a temperature sufficient to selectively lyse the platelets and red blood cells without damaging the white blood cells. The white blood cells of the lysed blood products are then contacted with a suitable dye to stain the white blood cells and the number of stained white blood cells is measured. The lysing reagent is free of harsh organic solvents which can damage the plastic components of automated clinical analyzers.

The present invention provides therapeutic mammalian cells which synthesize and express SS hemoglobin and a tumoricidal transgene. They are produced by transduction of SS erythroid progenitors / erythroblasts using viral vectors comprising a tumoricidal transgene operatively linked to the coding region of SS β-globin promoter / enhancer. Such transduced SS erythroid cells differentiate into mature SSRBCs that exhibit sustained synthesis and expression of SS hemoglobin, a tumoricidal protein(s). Both mature and progenitor SS-cells carrying tumoricidal transgene(s) are capable of selectively localizing in tumor microenvironment, occluding tumor microvessels and inducing a tumoricidal response.

The invention discloses a method for inducing direct transdifferentiation of mesenchymal stem cells into erythroblasts, and discloses a method for rapidly inducing the direct transdifferentiation of the mesenchymal stem cells (MSCs) into the erythroblasts (EBCs). The method comprises the following steps: S1, separating and purifying the homogeneous umbilical cord mesenchymal stem cells; S2, selecting small RNA molecules applicable to the epigenetic regulation of the stem cells; S3, preparing small nucleic acid polypeptide nanoparticles and conducting cell transfection; and S4, conducting amplification culture on the transfected erythroblasts. According to the method disclosed by the invention, neonatal umbilical cord mesenchymal stem cells, which are easily acquired, sufficient in source, free from ethical and moral problems and safe and effective, are taken as an induced object and six small RNAs molecules are transfected, so that the large-scale and rapid direct transdifferentiation of the stem cells into the erythroblasts under the action of a special serum-free medium is achieved; and the technical difficulties in the prior art of the artificial hematopoietic technology on the limitation of the source and the quantity as well as directional differentiation of the erythroblasts are solved.

The invention belongs to the field of cell biology detection, and particularly relates to a separation and purification method for foetus erythroblast in maternal blood and a separator. The method comprises the steps of (1) culturing a CD71 positive cell line (K562) and detecting antigen binding capacity and specificity; (2) establishing a catching system by a CTCBIOPSY-detection system and K562 cells and optimizing capturing conditions by cord blood; and (3) separating the collected foetus erythroblast: enlarging the diameter of target cells by microspheres coupled with a CD71 polyclonal antibody, and through the CTCBIOPSY- detection system, separating the erythroblast in the maternal blood by a filtering principle. The method is high in capturing efficiency, low in cost, simsple in operation steps, short in consumption time and high in sensitivity.

An objective is to provide screening methods for substances having an activity of promoting maturation into hemocytes. It was revealed that in the maturation process of erythrocytes and platelets, RP S19 multimers are released to the outside of the cell and act on the C5a receptors of erythroblasts and megakaryocytes, thereby causing maturation of erythroblasts and megakaryocytes to erythrocytes and platelets. Based on such findings, the present invention provides methods of screening for substances having an activity of promoting maturation into hemocytes by targeting the RP S19 multimers and C5a receptors. The screening methods of the present invention can detect hemocyte maturation using an increase of the R4 RGS family as an indicator. Furthermore, the present invention provides agents for promoting hemocyte maturation, which comprise RP S19 multimers and a substance having an activity of promoting maturation into hemocytes as useful components. The present invention also provides methods for producing erythrocytes and platelets, which include the step of contacting RP S19 multimers and a substance having an activity of promoting maturation into hemocytes with erythroblasts or megakaryocytes.

There is provided a method for identifying at least one foetal erythroblast the method comprising: (a) detecting the expression of at least one foetal erythroblast specific marker selected from the group consisting of neutral amino acid transporter B (SLC1A5), solute carrier family 3 (activators of dibasic and neutral amino acid transport) member 2 isoform A (SLC3A2), Splice Isoform A of Chloride channel protein 6, Transferrin receptor protein 1, Splice Isoform 3 of Protein GPR107 precursor, Olfactory receptor 11H4, Splice Isoform 1 of Protein C9orf5, Cleft lip and palate transmembrane protein 1, BCG induced integral membrane protein BIGM103, Antibacterial protein FALL-39 precursor, CAAX prenyl protease 1 homolog, Splice Isoform 2 of Synaptophysin-like protein, Vitamin K epoxide reductase complex subunit 1-like protein 1, Splice Isoform 1 of Protein C20orf22 (ABHD12), Hypothetical protein DKFZp564K247 (Hypoxia induced gene 1 protein) (IPI Accession No. IPI00295621), Hypothetical protein DK-FZp586C1924 (IPI Accession No. IPI00031064), ALEX3 protein variant, Hypothetical protein MGC14288 (IPI Accession No. IPI00176708), protein with IPI Accession No. IPI00639803 and protein with IPI Accession No. IPI00646289, wherein detection of the marker indicates the presence of the foetal erythroblast.

Techniques are provided for detecting hematological disorders using cell-free DNA in a blood sample, e.g., using plasma or serum. For example, an assay can target one or more differentially-methylated regions specific to a particular hematological cell lineage (e.g., erythroblasts). A methylation level can be quantified from the assay to determine an amount of methylated or unmethylated DNA fragments in a cell-free mixture of the blood sample. The methylation level can be compared to one or more cutoff values, e.g., that correspond to a normal range for the particular hematological cell lineage as part of determining a level of a hematological disorder.

The invention discloses a high-throughput rapid classification and analysis method for blood cells suitable for common fish such as crucian carp and carp. Aiming at the problem that nucleated red blood cells that cannot be removed interfere with white blood cell classification and counting in terms of quantity and shape, an automated measurement and analysis strategy combining fluorescent dyes and multi-channel images is used to separate the interference The red blood cell samples are separated from the white blood cells, and the range of different white blood cell groups that undertake important immune functions such as lymphocytes, monocytes, and granulocytes is divided, and automatic analysis is completed. The difficulty of effectively removing nucleated red blood cells has long plagued the automatic analysis of blood images of many types of animals. This invention combines fluorescent dyes with image analysis, and uses the high-throughput and high-sensitivity measurement modules of the multi-dimensional panoramic analyzer to accurately identify red blood cell groups. Eliminating its interference provides important technical support for focused analysis of leukocyte groups and has broad application prospects.

The invention discloses a method for separating and identifying nucleated erythrocytes in blood, which comprises the following steps: (1) preparing reagents; (2) preparing anti-degumming glass slides; Operation method to obtain nucleated red blood cells in the blood. The method for separating and identifying nucleated red blood cells in blood of the present invention can effectively identify nucleated red blood cells, all 40 samples can detect nucleated red blood cells, the detection rate is 100%, and 2×106 to 6×106 nucleated red blood cells were found in each case ml. The present invention is easy to identify nucleated red blood cells after benzidine staining, and can quickly obtain single nucleated red blood cells by skilled micromanipulation, laying a good preliminary foundation for applying single nucleated red blood cells to gene analysis.

The present invention provides a human cell population that can self-renew extensively and yet retain the capacity to differentiate into red blood cells (RBCs). These cells are referred to as extensively self-renewing erythroblasts (ESREs). The cells of the invention serve among other things as a renewable source of transfusable RBCs.