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

Method for culturing peripheral blood DC cells loaded with tumor cell exosome

A technology of tumor cells and culture methods, which is applied in the field of peripheral blood DC cell culture loaded with tumor cell exosomes, can solve the problems of inability to play a specific killing effect and complex protein components, and achieve the promotion of DC cell maturation and high content , high yield effect

Inactive Publication Date: 2019-03-19
中生康元生物科技(北京)有限公司
View PDF2 Cites 2 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The extracted protein components are complex and contain tumor intracellular antigens. After DCs are activated, DCs enter the body and present them to T cells. However, the antigens are expressed in the tumor cells because they cannot play a specific killing role.

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 culturing peripheral blood DC cells loaded with tumor cell exosome
  • Method for culturing peripheral blood DC cells loaded with tumor cell exosome
  • Method for culturing peripheral blood DC cells loaded with tumor cell exosome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0078] Preparation of tumor cell exosomes

[0079] 1. Medium

[0080] The components of the normal complete medium for tumor cells are: RPMI-1640 culture medium and FBS, and the amount of FBS added is 10% of the volume of the RPMI-1640 culture medium.

[0081] The components of the complete culture medium for exosomes are: phenol red-free RPMI-1640 culture medium and exosome-free FBS, and the amount of exosome-free FBS is 10% of the volume of the phenol red-free RPMI-1640 culture medium.

[0082] 2. Preparation of exosome-free FBS

[0083] Pour FBS into special ultracentrifuge tubes under sterile conditions, 38.6mL per tube, 4 tubes, after confirming the balance, put it into an ultracentrifuge at 120000g, and centrifuge overnight.

[0084] The next day, carefully draw the upper layer of clarified serum into a new sterile centrifuge tube, filter it with a 0.22 μm filter membrane into another centrifuge tube to ensure the sterility of the serum, seal it, and set it aside.

[...

Embodiment 2

[0097] A method for culturing peripheral blood DC cells loaded with tumor cell exosomes, comprising the following content:

[0098] 1. Peripheral blood DC cell culture medium

[0099] 1) DC medium

[0100] Based on the AIM-V medium, the following components were added according to the following contents: autologous plasma volume percentage was 1%±0.2%, GM-CSF concentration was 50ng / mL, and IL-4 concentration was 50ng / mL.

[0101] 2) DC maturation medium

[0102] Based on the AIM-V medium, add the following components according to the following content: the volume percentage of autologous plasma is 1%±0.2%, the concentration of GM-CSF is 50ng / mL, the concentration of IL-4 is 50ng / mL, and the concentration of TNF-α is 10ng / mL mL, PGE-2 concentration 1μg / mL.

[0103] 2. Peripheral blood DC cell culture method

[0104] Take 1 T175 culture flask, add the obtained PBMC cells into the culture flask, the number of cells in each flask is 1.5×10 8 , add 50 mL of AIM-V medium, add 5...

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 field of DC cell culture, and particularly relates to a method for culturing peripheral blood DC cells loaded with tumor cell exosome. The method comprises the following steps: performing adherent culture on a PBMC cell, removing culture solution and washing the cell; adding a DC culture medium, and culturing for 64-80 hours; patting a culture bottle to float the adherent cell, transferring the cell into a culture container coated with autologous plasma; and adding the DC mature culture medium, and culturing for 45-55 hours to obtain the peripheral blood DC cell loaded with tumor cell exosome. According to the method for culturing the peripheral blood DC cell loaded with tumor cell exosome, the tumor cell exosome can be used for promoting mature of DC cells, the functional phenotypes CD11c / CD83, CD11c / CD86 and HLA-DR / CD40 of the DC cell are superior to those of DC without exosome loading.

Description

technical field [0001] The present invention relates to the field of DC cell culture, in particular to a peripheral blood DC cell culture method loaded with tumor cell exosomes. Background technique [0002] Dendritic cells (DCs) are the most effective antigen-presenting cells in the body, express B7, MHC-II molecules, ICAM-1 and other co-stimulatory molecules on the surface, activate helper T cells, and prompt them to secrete IL- 2 and other cytokines transmit specific antigen signals and finally activate cytotoxic T cells. Therefore, DC cells play a key role in initiating a series of immune responses in the body, and are of great significance in the immune function of the body and the immune management of tumors. [0003] Most DC cells originate from the bone marrow, enter the peripheral blood from the bone marrow, and then distribute to various tissues throughout the body. DCs are widely distributed in all organs of the body except the brain, but the number is very small,...

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 Applications(China)
IPC IPC(8): C12N5/0784C12N5/09
Inventor 程旭东
Owner 中生康元生物科技(北京)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