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

Enhanced cik cell preparation method and cell preparation

A cell preparation, enhanced technology, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problem that immune cells cannot achieve the effect of killing tumors, CIK cell proliferation times and cytotoxic activity are not ideal, etc. problem, to achieve the effect of increasing the killing effect and increasing the toxicity

Active Publication Date: 2017-05-17
ANDERSON CELL BIOLOGY HUBEI
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Therefore, the effect of killing tumors cannot be achieved by relying on one's own immune cells, and the absolute number of such cells can only be increased by culturing in vitro
[0004] CIK cells can proliferate in large quantities through in vitro induction. The general CIK preparation method is to treat isolated peripheral blood mononuclear cells (PBMC) with IFN-γ for 24 hours, and then add CD3mAb, IL-1α and IL-2 factors to stimulate and induce , and finally obtain a certain number of CIK cells, but the multiplier and cytotoxic activity of the finally obtained CIK cells are not ideal

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
  • Enhanced cik cell preparation method and cell preparation
  • Enhanced cik cell preparation method and cell preparation
  • Enhanced cik cell preparation method and cell preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] A method for preparing enhanced CIK cells, comprising the following steps:

[0041] 1) Collect the patient’s anticoagulant blood under sterile conditions, centrifuge at 1500rpm / min for 6 minutes to absorb the upper plasma, inactivate at 56°C for 30 minutes and then centrifuge for use. Dilute the blood cell pellet with compound normal saline with the same volume as the blood cell pellet, with a specific gravity of 1.077 Human lymphocyte separation solution and diluted blood were added to the centrifuge tube at a ratio of 1:1.5, centrifuged at 2000rpm / min for 15 minutes, carefully extracted the white blood cell layer in the middle, washed twice with compound normal saline, and obtained peripheral blood mononuclear cells after low-speed centrifugation (PBMC);

[0042] 2) Peripheral blood mononuclear cells were isolated from peripheral blood collected from patients; cultured in blood-free medium to obtain 1×10 6 nucleated cells / ml, add IFN-γ, phytohemagglutinin and PGE2, a...

Embodiment 2

[0062] A method for preparing enhanced CIK cells, comprising the following steps:

[0063] 1) Collect the patient’s anticoagulant blood under sterile conditions, centrifuge at 1500rpm / min for 6 minutes to absorb the upper plasma, inactivate at 56°C for 30 minutes and then centrifuge for use. Dilute the blood cell pellet with compound normal saline with the same volume as the blood cell pellet, with a specific gravity of 1.077 Human lymphocyte separation solution and diluted blood were added to the centrifuge tube at a ratio of 1:1.5, centrifuged at 2000rpm / min for 15 minutes, carefully extracted the white blood cell layer in the middle, washed twice with compound normal saline, and obtained peripheral blood mononuclear cells after low-speed centrifugation (PBMC);

[0064] 2) Peripheral blood mononuclear cells were isolated from peripheral blood collected from patients; cultured in blood-free medium to obtain 1×10 6 nucleated cells / ml, add IFN-γ, phytohemagglutinin and PGE2, a...

Embodiment 3

[0069] A method for preparing enhanced CIK cells, comprising the following steps:

[0070] 1) Collect the patient’s anticoagulant blood under sterile conditions, centrifuge at 1500rpm / min for 6 minutes to absorb the upper plasma, inactivate at 56°C for 30 minutes and then centrifuge for use. Dilute the blood cell pellet with compound normal saline with the same volume as the blood cell pellet, with a specific gravity of 1.077 Human lymphocyte separation solution and diluted blood were added to the centrifuge tube at a ratio of 1:1.5, centrifuged at 2000rpm / min for 15 minutes, carefully extracted the white blood cell layer in the middle, washed twice with compound normal saline, and obtained peripheral blood mononuclear cells after low-speed centrifugation (PBMC);

[0071] 2) Peripheral blood mononuclear cells were isolated from peripheral blood collected from patients; cultured in blood-free medium to obtain 1×10 6 nucleated cells / ml, add IFN-γ, phytohemagglutinin and PGE2, and...

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 discloses a preparation method of an enhanced CIK cell, and a cell preparation. The method comprises the following steps: separating to obtain a peripheral blood mononuclear cell from peripheral blood collected from a sufferer; putting the peripheral blood mononuclear cells into a blood-free medium to culture, so as to obtain karyocyte of which the concentration is (1-3)*10<6> / ml; adding IFN (interferon)-gamma, phytohemagglutinin and PGE2 (Phenyl Glycidyl Ether), transferring to a culture bottle or a culture bag to culture for 24 hours, and then adding a CD3 monoclonal antibody, IL-2 (interleukin-2), IL-1alpha, IL-4 and GM-CSF (granulocyte-macrophage colony-stimulating factor); continuing to culture for 3-5 hours, and then controlling the cell density at (1-6)*10<6> / ml; and centrifugally colleting to obtain the enhanced CIK cell after continuously culturing for the 14th to 21st days. The enhanced CIK cell preparation comprises the enhanced CIK cell, human serum albumin, IL-2, amino acids, vitamins and inorganic salts. Compared with a preparation in the patient, the CIK preparation has the advantages that the preservation time of the CIK preparation is obviously prolonged, and is prolonged to 36 hours from 6 hours at room temperature. The CIK preparation is higher in reliability in clinical application, and the safety is effectively ensured.

Description

technical field [0001] The invention relates to the field of in vitro culture of immune cells, in particular to a method for preparing enhanced CIK cells and a cell preparation. Background technique [0002] Adoptive cellular immunotherapy refers to a treatment method that directly kills tumor cells or viruses by infusing self or allogeneic specific and non-specific anti-tumor immune effector cells. Adoptive cellular immunotherapy has entered clinical application and has become the fourth major tumor treatment method after surgery, radiotherapy and chemotherapy. The types of such immune cells include lymphokine-activated killer cells (LAK), tumor-infiltrating lymphocytes (TIL), cytotoxic lymphocytes (CTL), etc. Among many immune cells, CIK cells have tumoricidal activity High, broad tumor killing spectrum, equally sensitive to a variety of drug-resistant cells, and less toxic to normal bone marrow hematopoietic premise cells, etc., are widely used in clinical practice. [...

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): C12N5/0783A61K35/17A61P35/00A61K35/14
Inventor 许亮艾伟石纪刚柳海洋张辉
Owner ANDERSON CELL BIOLOGY HUBEI
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