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

msc-tnf α-AB stem cell and its preparation method and application

A technology of MSC-TNF and stem cells, which is applied in the field of MSC-TNFα-AB stem cells and its preparation, can solve the problems of high recurrence rate, large side effects, and insignificant therapeutic effect, and achieve the effect of avoiding allergic reactions

Active Publication Date: 2021-02-09
连祺周
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0006] At present, there is no report on the combination of MSC and TNF α antibody at home and abroad. However, the traditional treatment of osteoarthritis has a high recurrence rate, large side effects and ineffective therapeutic effect. Therefore, the effective combination of MSC and TNF α antibody can make TNF α antibody can be activated by the inflammatory factor TNF α in MSC, which is expected to be the key point of gene therapy for osteoarthritis

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
  • msc-tnf α-AB stem cell and its preparation method and application
  • msc-tnf α-AB stem cell and its preparation method and application
  • msc-tnf α-AB stem cell and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0052] Example 1. Preparation of Osteoarthritis Inflammatory Infiltration Mouse Model

[0053] Dissolve the purified natural collagen in 11M acetic acid to make a solution with a concentration of 1mg / mL, and mix it with an equal amount of Freund’s adjuvant or incomplete adjuvant to form a stable emulsion; use 1mL of the emulsion to inject 4 to 6 sites Intradermally on the back of the mouse, small ulcers will appear at the injection site (heal after 7-10 days); 21 days later, dissolve 15 mg of collagen in 15 mL of 1M acetic acid and inject it into the abdominal cavity of the mouse for another immunization.

[0054] 14 to 60 days after the initial injection, joint redness and swelling appear, usually in the distal joints, and the rear foot ankle joint is the most common site of accumulation. The pathological changes in the acute phase can be divided into two phases. After 10 days of immunization, there are only some fibrous deposits in the joints and no changes in the synoviocyt...

Embodiment 2

[0055] Example 2. Design and synthesis of TNF α-AB antibody fragments, lentivirus preparation, MSC infection

[0056] The packaging plasmid required for this experiment was purchased from addgene company. The plasmid map is shown in Figure 3, and Figure 3 includes Figure 3A , Figure 3B and Figure 3C ; The shuttle plasmid pGFP was purchased from Guangzhou Funeng Gene Co., Ltd., see the plasmid map Figure 4 ; The information of the above plasmids is shown in Table 1; lentiviral packaging cells 293FT were purchased from Invitrogen (catalog number R700-07); MSCs were extracted from the bone marrow of C57 / BL6 mice; C57 / BL6 mice were purchased from the Experimental Animal Center of the University of Hong Kong.

[0057] The target fragment sequence TNF α-AB required in this experiment is shown in SEQ ID NO:1.

[0058] The reagents required for this experiment are listed in Table 1.

[0059] Table 1 Lentiviral packaging plasmid and shuttle plasmid information

[0060]

[0...

Embodiment 3

[0101] Example 3. Detection of the constructed new stem cell line MSC-TNF α-AB can be specifically regulated and initiated by the inflammatory factor TNF α

[0102] This example is to detect the expression of TNF α-AB after adding the inflammatory factor TNF α in MSC cell culture in vitro.

[0103] 3.1 MSC-TNF α-AB cell culture:

[0104] MSC-TNF α-AB cell culture medium is: in DMEF / F12 medium containing 10% fetal bovine serum (FBS), 100U / mL penicillin streptomycin, 1% non-essential amino acids, 0.1% β-mercaptoethanol, 5ng / mL of EGF, 5ng / mL of FGF.

[0105] 1) Before aseptic culture, autoclave the materials required in the laboratory; irradiate with ultraviolet light for 30 minutes; wear special lab coats and gloves between cells, and wipe hands with 75% alcohol; observe the state of cells under a microscope, and the cell density is confluent If it reaches more than 70%, it needs to be passed down, turn off the ultraviolet lamp, turn on the fan, and at the same time wipe the...

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 relates to the technical field of stem cell therapy, in particular to a MSC-TNFα-AB stem cell and its preparation method and application. MSC‑TNFα‑AB stem cells are recombinant mesenchymal stem cells that integrate TNFα antibody genes. The present invention also provides a promoter capable of positively activating and expressing TNFα-AB in MSC-TNFα-AB stem cells in the presence of inflammatory factor TNFα. The present invention overexpresses the TNFαAB gene in mesenchymal stem cells through the in vitro lentivirus method, and the constructed MSC TNFα‑AB stem cells will be activated by TNFα inflammatory factors in an inflammatory environment, and more stably bind to the inflammatory site, thereby strengthening the anti-inflammatory effect of MSCs. Inflammation and immune regulation ability, effectively relieve the inflammatory response of osteoarthritis.

Description

technical field [0001] The invention relates to the technical field of stem cell therapy, in particular to a MSC-TNFα-AB stem cell and its preparation method and application. Background technique [0002] Osteoarthritis (OA), also known as degenerative osteoarthritis, is the degeneration of articular cartilage and the formation of bony spurs at the edge of joints. As time goes on, the development of osteoarthritis is getting faster and faster. Inflammation can promote the degeneration of cartilage, and cartilage degeneration can stimulate the development of inflammation. Inflammation is mainly through the destruction of cell matrix and chondrocytes through inflammatory factors, and the main signaling pathways include MAPK, nuclear transcription factor signaling pathway, Wnt signaling pathway, etc. The methods of treating osteoarthritis include drug therapy, non-drug therapy such as physical therapy, physical exercise, weight control, orthopedics and surgery. The current mai...

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): C12N5/10C12N15/113C12N15/867A61K39/395A61K35/28A61P19/02A61P29/00
CPCA61K35/28A61K39/395A61K48/005A61K2039/5156C07K16/241C12N15/86C12N2740/15043A61K2300/00
Inventor 连祺周
Owner 连祺周
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