Application of histone methyltransferase inhibitor to preparation of product for promoting megakaryocyte proliferation or blood platelet formation

A methyltransferase and megakaryocyte technology, applied in the field of preparation of products that promote megakaryocyte proliferation or platelet production, histone methyltransferase inhibitors, can solve problems such as low platelet levels, and achieve low application cost and platelet production Efficient, maneuverable and reproducible results

Active Publication Date: 2019-12-20
INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] One aspect of the present invention is aimed at the lack of compounds and methods that simultaneously promote cell proliferation, megakaryotic differentiation and platelet production in the prior art, and the low l

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of histone methyltransferase inhibitor to preparation of product for promoting megakaryocyte proliferation or blood platelet formation
  • Application of histone methyltransferase inhibitor to preparation of product for promoting megakaryocyte proliferation or blood platelet formation
  • Application of histone methyltransferase inhibitor to preparation of product for promoting megakaryocyte proliferation or blood platelet formation

Examples

Experimental program
Comparison scheme
Effect test

Example Embodiment

[0060] Example 1:

[0061] 1) Cord blood CD34 + Cell separation

[0062] a. Pour the cord blood into a plasma bottle or 50ml centrifuge tube, follow V Cord blood :V Sorting buffer =1:1 Add the sorting buffer (the formula is as follows), then add the hydroxyethyl starch of 1 / 4 of the above mixing volume and mix well, and let it stand at room temperature for 45 minutes;

[0063] Sorting buffer formula: phosphate buffered saline (PBS), 0.5% bovine serum albumin (BSA), 0.4% 500mM ethylenediaminetetraacetic acid (EDTA), 1% penicillin streptomycin mixture (P / S)

[0064] b. Transfer the supernatant to a 50ml centrifuge tube, centrifuge at 1500rpm for 10min at room temperature; discard the supernatant, add sorting buffer to resuspend the cells; take a 15ml centrifuge tube, add 4.5ml Ficoll separation solution (V Ficoll :V Sorting buffer =1:1), slowly add the above-mentioned resuspension to the Ficoll separation liquid to avoid breaking the liquid level, 1800rpm, 15min (the centrifuge is s...

Example Embodiment

[0075] Experimental example 1: Cell proliferation detection

[0076] Count the cells in each stage in Example 1, and set up different control groups.

[0077] a. The control group (DMSO) and the experimental group (EHMTi) were cultured under the same conditions, and the cells were counted every three days to detect the effect of small molecule compounds on cell proliferation.

[0078] b. The result is figure 1 As shown, compared with the DMSO control group, adding histone methyltransferase inhibitor (A366) treatment on day 0-3 or day 3-6 of culture can significantly promote cell proliferation and increase the cell expansion ratio by 15 times the above.

[0079] figure 1 The total number of cells in the control group (DMSO group) showed a downward trend on the 9th-18th day of mid-megakaryotic differentiation; after treatment with histone methyltransferase inhibitors, the total number of cells still showed an upward trend on the 9th-12th day of megakaryocyte differentiation, and megakar...

Example Embodiment

[0080] Experimental example 2: Flow cytometric detection of megakaryocytes

[0081] The megakaryocytes in Example 1 were detected.

[0082] a. Collect about 1×10 on the 3rd, 6th, 9th, and 12th day of megakaryocyte differentiation 5 The cell suspension was centrifuged at 300 g for 5 min, and the cell pellet was resuspended with 100 μL 0.2% BSA.

[0083] b. Add 1μL of anti-CD41a-APC (BD) and 1μL of anti-CD42b-PE (BD) flow cytometry antibody to each group. After incubating for 30 minutes in the dark, flow cytometry (FACS Canto II; BD Biosciences) detects CD41a + CD42b + Proportion of megakaryocytes.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

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.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the use of histone methyltransferase inhibitors in the preparation of products for promoting megakaryocyte proliferation or platelet production. Background technique [0002] Platelets are one of the formed components of blood, and play an important role in the process of hemostasis and thrombus in the body through functions such as adhesion, aggregation, and particle release. Clinically, various etiologies such as malignant tumors of the hematopoietic system, aplastic anemia, malignant tumors, hematopoietic stem cell transplantation therapy, and radiotherapy and chemotherapy may lead to thrombocytopenia or dysfunction, resulting in varying degrees of bleeding in the body, and even endangering the body in severe cases. life. Donor-derived platelet concentrate transfusion is currently the only clinical treatment. According to statistics, the number of patients who need platelet tran...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C12N5/0786C12N5/078A61K35/19A61K35/28A61P7/06A61P7/04A61P1/16
CPCA61K35/19A61K35/28A61P1/16A61P7/04A61P7/06C12N5/0644C12N5/0645C12N2500/90C12N2501/125C12N2501/145C12N2501/2303C12N2501/72C12N2506/11
Inventor 周家喜刘懿莹刘翠翠苏培王洪涛
Owner INST OF HEMATOLOGY & BLOOD DISEASES HOSPITAL CHINESE ACADEMY OF MEDICAL SCI & PEKING UNION MEDICAL COLLEGE
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap