Ex vivo amplification culture system of umbilical cord hematopoietic stem cells and ex vivo amplification method for umbilical cord hematopoietic stem cells

A technology for hematopoietic stem cells and a culture system, which is applied in the field of umbilical cord hematopoietic stem cells in vitro expansion and culture system, can solve the problem of difficulty in meeting the large demand of HSCs and the like

Inactive Publication Date: 2020-03-20
李陶 +1
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In view of this, the present invention provides an umbilical cord hematopoietic stem cell in vitro expansion culture system and an umbilical cord hematopoietic stem cell in vitro expansion method, which are

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
  • Ex vivo amplification culture system of umbilical cord hematopoietic stem cells and ex vivo amplification method for umbilical cord hematopoietic stem cells
  • Ex vivo amplification culture system of umbilical cord hematopoietic stem cells and ex vivo amplification method for umbilical cord hematopoietic stem cells
  • Ex vivo amplification culture system of umbilical cord hematopoietic stem cells and ex vivo amplification method for umbilical cord hematopoietic stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] This embodiment carries out the preparation of umbilical cord blood mononuclear cells, comprising the following steps:

[0028] Add heparin sodium (20mg / L) in a 100ml glass bottle for anticoagulation, and collect the cord blood of fresh full-term pregnancy. Among them, the collection of cord blood excludes infectious diseases and various family genetic diseases, and the cord blood collection is strictly aseptic operate. The average volume of the collected cord blood is 50ml-100ml, and the cord blood is separated within 4 hours after collection. When separating, divide the umbilical cord blood into several sterile centrifuge tubes and centrifuge at 1500rpm for 10min. After centrifugation, transfer the upper layer of plasma to a new centrifuge tube, inactivate at 56°C for 30 minutes, let stand at -20°C for 10 minutes, and centrifuge at 2000rpm for 10 minutes, then take the upper layer of plasma for use. After centrifugation at 1500rpm for 10min, blood cells were obtaine...

Embodiment 2

[0030] In this example, the content of CD34+ cells was detected on the umbilical cord blood mononuclear cells prepared in Example 1. The results were detected by flow cytometry figure 2 As shown, the results showed that the CD34+ cell content in cord blood mononuclear cells was 1.25%.

Embodiment 3

[0032] In this embodiment, the umbilical cord blood mononuclear cells prepared in Example 1 are divided into eight groups, corresponding to the following different experimental groups, cultured in different culture systems, cultured in 24-well plates, and the volume of the culture system is 0.5 ml per well, The cell seeding density was 2×10 6 cells / mL, half the volume was changed every two days. Wherein, the plasma in the culture system is the plasma prepared in Example 1. Observe the cell growth density under an inverted microscope at the 1st, 2nd, 3rd, and 4th week, and count the suspension cells at the same time. The results are as follows: figure 2 As shown, compared with control group 1, the proliferation rate of cord blood mononuclear cells in experimental group 1 to experimental group 7 was faster, indicating that G1 / S-specific cyclin-D1 (cyclins D1) and bone morphogenetic protein 4 ( Bone morphogenetic protein 4, BMP4) can promote the proliferation rate of umbilical...

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 belongs to the technical field of stem cells, and especially relates to an ex vivo amplification culture system of umbilical cord hematopoietic stem cells and an ex vivo amplification method for the umbilical cord hematopoietic stem cells. The provided system includes G1/S-specific cyclin-D1 and bone morphogenetic protein 4. Through the indication of results, the ex vivo amplification efficiency of the umbilical cord hematopoietic stem cells can be obviously enhanced and the dryness of the umbilical cord hematopoietic stem cells can be maintained by adding the G1/S-specific cyclin-D1 and bone morphogenetic protein 4 into the ex vivo amplification culture system of umbilical cord hematopoietic stem cells.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to an in vitro expansion culture system of umbilical cord hematopoietic stem cells and an in vitro expansion method for umbilical cord hematopoietic stem cells. Background technique [0002] Hematopoietic stem cells (HSCs), as one of the blood components, are the initial cells that generate various blood cells, also known as hematopoietic pluripotent stem cells, which exist in bone marrow, embryonic liver, peripheral blood and umbilical cord blood. HSCs have both a high capacity for self-renewal and the ability to further differentiate into hematological progenitor cells. At present, HSCs have been widely used in hematopoietic stem cell transplantation clinically, and have broad application prospects in gene therapy, immunotherapy and preparation of mature blood cell products. Bone marrow has always been the main source of HSCs, but the difficulty in finding a suitab...

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): C12N5/0789
CPCC12N5/0647C12N2500/32C12N2500/90C12N2501/125C12N2501/14C12N2501/155C12N2501/2303C12N2501/2306C12N2501/405
Inventor 李陶
Owner 李陶
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