Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

A production method for promoting cytokine production in stem cells

A cytokine and stem cell technology, applied in the field of promoting the production of stem cell cytokines, can solve the problems of not verifying the adipogenic differentiation of bone marrow mesenchymal stem cells, and not involving the production and enrichment of cytokines, and achieve the effect of promoting high secretion

Active Publication Date: 2022-07-01
上海天安智谷干细胞科技集团有限公司
View PDF4 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in the prior art, it has not been verified to promote the adipogenic differentiation of bone marrow mesenchymal stem cells after rapid proliferation, detect the changes of corresponding cytokines, and there is no research involving the production and enrichment of corresponding cytokines.

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
  • A production method for promoting cytokine production in stem cells
  • A production method for promoting cytokine production in stem cells
  • A production method for promoting cytokine production in stem cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Example 1 Preparation of monoclonal antibodies specific for PTN

[0020] The optimized PTN highly immunogenic epitope peptide sequence rtgaeckqtmktqrckipcnwkkqfgaeckyqfqawgecdlntalktrtg was synthesized and used for later use. Animal Immunization: Four 7-week-old female Balb / c mice were taken, and the antigen was fully emulsified with an equal volume of Freund's adjuvant at a dose of 50 μg / mice, and then injected subcutaneously at multiple points in the abdomen, and boosted immunization every 2 weeks. Seven days after the third immunization, the immunized mouse serum was collected by enucleating the eyeball method, which was the immune serum (positive serum). Another free mouse serum was collected as negative serum. On the 7th day after 3 times of immunization, blood was collected from the tail vein, and the serum titer of mice was determined by ELISA. 10μg / mL, coated at 4°C for 12h; 50mg / L nonfat dry milk was blocked at 37°C for 1h; mouse serum was diluted with PBST, ...

Embodiment 25

[0024] Example 2 Subtype identification of 5H2 monoclonal antibody

[0025]Sigma's Mouse Monoelonal Antibody Isotyping Kit was used for identification. The hybridoma cells and mouse ascites were taken and diluted 1:10 with PBS with pH 7.2; 200 μL of the dilution was added to the test tube, kept at room temperature for 30 s, the Isotrip colloidal gold test paper was inserted into the bottom of the tube, taken out after 10 minutes, and the results were observed. The results showed that the antibody subtype of 5H2 monoclonal antibody was IgG1, and the light chain type was κ.

Embodiment 3

[0026] Example 3 Affinity identification and sequence identification of 5H2 monoclonal antibody

[0027] The affinity was determined by surface plasmon resonance (Surface Plasmon Resonance, SPR). For the immobilization of the capture molecule (anti-mouse Fc fragment capture antibody), the channel immobilized with the anti-mouse Fc fragment capture antibody is used as the detection channel, and the channel without the anti-mouse Fc fragment capture antibody is used as the control channel. The process is as follows: (1) Surface equilibration: HBS-EP buffer, equilibrate the chip surface at a flow rate of 15μl / min for 5min; (2) Surface activation: inject 'NHS+EDC' 1:1 mixture, activate the chip surface at a flow rate of 15μl / min for 7min; (3) Coupling Coupling antibody: inject goat anti-mouse Fc fragment capture antibody (diluted in 10 mM sodium acetate (pH 4.5) buffer), and couple at a flow rate of 15 μl / min for 6 min; control channel omit this step; (4) Surface blocking: inject ...

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
affinityaaaaaaaaaa
Login to View More

Abstract

The invention relates to a production method for promoting the production of stem cell cytokines. The method uses a monoclonal antibody that can specifically target PTN. The antibody can effectively induce adipogenic differentiation of bone marrow mesenchymal stem cells, and can efficiently differentiate after differentiation. To promote the high secretion of SCF and VEGF cytokines, the cytokines containing the high yields can be used for subsequent studies by isolating and purifying.

Description

technical field [0001] The present application relates to the biological field, and more particularly, to a production method for promoting the production of stem cell cytokines. Background technique [0002] Mesenchymal stem cells (MSCs) are important members of the stem cell family, which not only exist in bone marrow, fat, cord blood and peripheral blood, but also in perichondrium, periosteum, muscle and trabecular bone. At present, most researches are on bone marrow mesenchymal stem cells (BMSCs). Its basic characteristics are: self-renewal ability, can proliferate to form tissue and maintain its own number in vivo, and can grow clonally in vitro. The ability of proliferation and differentiation can differentiate into muscle, nerve, cartilage, bone, fat, tendon and ligament and other tissues under the induction of certain environment and specific factors. BMSCs have become the most commonly used seed cells for bone tissue engineering research due to their advantages of...

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): C12N5/0775C12N5/077C07K16/22C12P21/02C07K14/475C07K14/515
CPCC12N5/0663C12N5/0653C07K14/475C07K14/515C12P21/02C07K16/22C12N2506/1353C12N2501/33C12N2500/40C12N2501/999C12N2501/10C07K2317/56C07K2317/92
Inventor 张海涛李娜
Owner 上海天安智谷干细胞科技集团有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap 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