Eureka AIR delivers breakthrough ideas for toughest innovation challenges, trusted by R&D personnel around the world.

Method for amplifying candidate stem cell

A technology of hematopoietic stem cells and nuclear cells, applied in the direction of blood/immune system cells, animal cells, vertebrate cells, etc., can solve the problems of anti-leukemia and graft-versus-host effect changes, reduced proliferation ability, and failure to meet clinical use requirements, etc. , to reduce the differentiation of hematopoietic stem cells, promote the generation of stromal cells, and meet the requirements of clinical use

Inactive Publication Date: 2011-09-28
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the above methods have the following problems: (1) the scale of culture is difficult to scale up, which cannot meet the requirements of clinical use; (2) the co-cultivation of heterologous stromal cells and stromal cells and hematopoietic stem cells may have the problem of immune rejection, while the same The preparation process of source stromal cells is cumbersome and takes a long time; (3) Cytokines alone cannot maintain long-term hematopoiesis in vitro, and the amount of cytokines is large and expensive, which greatly limits the large-scale expansion of hematopoietic stem cells
However, when hematopoietic stem cells are separated and purified from mononuclear cells, part of them will inevitably be lost, and the highly purified hematopoietic stem cells will completely lose other auxiliary cells, which not only leads to a decrease in proliferation ability, but also causes graft-versus-leukemia and graft-versus-host effects. will change

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
  • Method for amplifying candidate stem cell
  • Method for amplifying candidate stem cell

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A method of expanding hematopoietic stem cells,

[0034] 1) Separation of mononuclear cells:

[0035] Obtaining nucleated cells from human umbilical cord blood: add methylcellulose solution with a weight concentration of 0.125% to human umbilical cord blood at a volume ratio of 4:1, let stand at room temperature, and settle red blood cells; take the supernatant at 1500rpm / min Centrifuge, the sediment is nucleated cells.

[0036] Density gradient centrifugation, the Ficoll lymphocyte separation solution with a density of 1.077 / ml was added to the nucleated cells in a volume ratio of 1:1, 1500rpm / mim, 30min, and the mononuclear cells in the middle layer were aspirated; it contained CD34 + The number of cells is 0.9% of the total number of cells; after washing with PBS (1500rpm / min, 5min), adjust the cell density for later use.

[0037] 2) Preparation of microencapsulated mononuclear cells:

[0038] A. Prepare sodium alginate with B-L-guluronic acid weight content of 31.2% and α-D-...

Embodiment 2

[0051] A method of expanding hematopoietic stem cells,

[0052] 1) Separation of mononuclear cells:

[0053] Density gradient centrifugation, the Ficoll lymphocyte separation solution with a density of 1.077 / ml was added to the human bone marrow fluid at a volume ratio of 1.5:1, 1500rpm / mim, 30min, and the mononuclear cells in the middle layer were aspirated; it contained CD34 + Number of cells 1.9% of the total number of cells; After washing with PBS (1500rpm / min, 5min), adjust the cell density for later use;

[0054] 2) Preparation of microencapsulated mononuclear cells:

[0055] A. Prepare sodium alginate with a weight content of 37% β-L-guluronic acid and 63% α-D-mannuronic acid into 1.0% w / v sodium alginate solution I;

[0056] Sodium alginate containing 62% by weight of β-L-guluronic acid and 38% by weight of α-D-mannuronic acid is prepared into 2.5% w / v sodium alginate solution II;

[0057] Mix sodium alginate solution I and sodium alginate solution II in a volume ratio of 1:1 to ...

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

PropertyMeasurementUnit
diameteraaaaaaaaaa
Login to View More

Abstract

The invention relates to a method for amplification of hemopoietic stem cells. The method comprises the steps of separation of mononuclear cells, preparation and culture of microencapsulated mononuclear cells, and amplification of hemopoietic stem cells. The method can realize the amplification of hemopoietic stem cell as well as carries out culture on a large scale, and can play a significant role in blood biology and clinic application and research.

Description

Technical field [0001] The invention relates to a method for amplifying hematopoietic stem cells under three-dimensional conditions. Specifically, it is to prepare microencapsulated mononuclear cells, and use the physical and chemical properties of microcapsules and microencapsulated materials to promote the production of stromal cells, and realize the three-dimensional co-culture of hematopoietic stem cells and homologous stromal cells. The stromal cells secrete extracellular matrix and cells at the same time. Factors, which together form a hematopoietic microenvironment in the microcapsules, can inhibit / delay the differentiation of hematopoietic stem cells and achieve effective expansion of hematopoietic stem cells with only a small amount of cytokines. Background technique [0002] Hematopoietic stem cells have a wide range of clinical applications. They have a wide range of applications in supportive therapy, immunotherapy, replacement therapy, and gene therapy for blood syst...

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
Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/06C12N5/08C12N5/0786C12N5/0789
Inventor 马小军李双月王为于炜婷
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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
Eureka Blog
Learn More
PatSnap group products

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

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com