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

Method for in-vitro amplification of NK cells

A technology for NK cells and in vitro expansion, which is applied in the field of NK cell culture and expansion, and can solve the problems of high cost and unsatisfactory purity

Inactive Publication Date: 2013-03-27
SHANGHAI CLAISON BIOTECH
View PDF4 Cites 34 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In order to solve the problems of unsatisfactory purity and high cost of NK cells after expansion in the prior art, the present invention provides a new method for in vitro expansion of NK cells, which can obtain a large number of high-purity, high-purity NK cells under in vitro culture conditions. NK cells with strong killing activity

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 in-vitro amplification of NK cells
  • Method for in-vitro amplification of NK cells
  • Method for in-vitro amplification of NK cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] 1. Isolation of peripheral blood mononuclear cells (PBMC) and culture of NK cells

[0027] Collect peripheral blood mononuclear cells (PBMC) from the patient through a blood cell separator, transfer the collected blood sample to a centrifuge tube; 700g, centrifuge for 10 minutes, and absorb the upper layer of plasma for later use; use 0.9% normal saline blood sample to restore the sample to its original volume, Mix well; slowly add the diluted blood to Ficoll, 900g, and centrifuge for 20min; absorb the milky white mononuclear cell layer at the interface of the separation solution; centrifuge and wash twice and count; resuspend PBMC with medium, and adjust the cell concentration to 1.5×10 6 / ml,

[0028] Transfer to a 75cm cell line pre-coated with CD3McAb and CD226McAb 2 In the culture flask, add 500IU / ml IL-2 and 10ng / ml IL-18, culture for 72 hours,

[0029] The cells were collected by centrifugation, counted, and mixed with the same number of K562 cells killed by 1...

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
concentrationaaaaaaaaaa
Login to View More

Abstract

The invention relates to a method for in-vitro amplification of NK cells, and in particular relates to a method for massive in-vitro amplification of NK cells, wherein the method comprises the following steps of: a, inoculating a peripheral blood mononuclear cell in a CD3McAb and CD226McAb pre-coated culture bottle for coculture; b, adding 1L-2 and 1L-18, coculturing for 72hours to stimulate amplification of NK cells; c, transferring the NK cells, K562 cells after lethal treatment and a serum-free medium containing 1L-2 and 1L-18 in a cell culture bag for coculture; and d, collecting the NK cells. According to the method for in-vitro amplification of the NK cells, two antibodies CD3McAb and CD226McAb are simultaneously coated, so the cell factor synthesis and ADCC effect are promoted, and killing toxicity of the NK cells is remarkably improved; the activation and amplification on the NK cells are achieved just by the 1L-2 and 1L-18 cell factors, so the amplification multiple and cell toxicity of the NK cells are guaranteed, and the cost of cell culture is reduced.

Description

[technical field] [0001] The invention relates to a method for cultivating and expanding NK cells, in particular to a method for massively expanding NK cells in vitro. [Background technique] [0002] Natural killer cells (NK) are cytotoxic lymphocytes belonging to the lymphocyte lineage that contain perforin and granzyme granules. Its recognition of target cells is not restricted by MHC, it can directly kill tumor cells without prior sensitization, and can also secrete cytokines to regulate the functions of other immune cells. It is the main bearer of the body's natural immunity and the core of acquired cellular immunity. Regulatory cells play an important role in tumor immunity, anti-viral infection and removal of non-self cells. Studies have proved that natural killer cells are not only an important component of the natural immune system, but also have some characteristics of acquired immune cells. In the immune system, NK cells respond faster than T cells or B cells, an...

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): C12N5/0783
Inventor 叶永清杨雪晶苏国新
Owner SHANGHAI CLAISON BIOTECH
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