Method for in-vitro culture of killer T cells

A technology of cells and culture medium, applied in the field of culturing killer T cells in vitro, can solve the problems of many operation steps, high reagent cost, combined stimulation, etc., and achieve the effect of simple operation steps, low reagent cost and good vitality

Active Publication Date: 2013-04-03
JILIN TUO HUA BIOTECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CTL, as specific cytotoxic T cells, needs to be induced by tumor tissue during culture, which limits the culture
CIK cell culture requires combined stimulation of 4 factors, many steps and high cost of reagents

Method used

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  • Method for in-vitro culture of killer T cells
  • Method for in-vitro culture of killer T cells
  • Method for in-vitro culture of killer T cells

Examples

Experimental program
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Embodiment 1

[0046] 1. Raw materials: 50ml of sterile human whole blood was obtained and sent to the central laboratory of our hospital for extraction and cultivation.

[0047] 2. Reagents, consumables and instruments

[0048] 2.1 Main reagents and consumables: RPMI 1640 serum-free lymphocyte culture medium (purchased from Tianjin Haoyang Biotechnology Company, containing stable glutamine, phenol red-free, and various serum substitutes), recombinant human cytokine IL- 2 (Shanghai Huaxin Biotech Co., Ltd.), anti-CD3 monoclonal antibody (purchased from Peprotech, USA), 0.01M, pH7.2 phosphate buffered saline (PBS), Folium Folium (FS, Chinese herbal medicine Folium Folium Produced in Hebei, purchased by medicinal materials company, Brassicaceae), cell culture flasks, centrifuge tubes and pipettes were purchased from Corning Company, USA.

[0049] Preparation of Folium Folium decoction: 50g of Folium Folium dried medicinal material, soaked in cold water for 20-30 minutes, concentrated the deco...

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Abstract

The invention relates to a method for in-vitro culture of killer T cells. Specifically, IL-2 and an anti-CD3 monoclonal antibody are combined with indigowoad leaf decoction for costimulation of lymphocytes taken from 50ml of in-vitro blood so as to get the T cells with cytotoxin activity by induction; and the T cells with the cytotoxin activity, which are obtained by an induction culture method, have the advantages of large proliferation number and good vitality, and the number of the CD8-positive T cells can be increased from 20%-30% to 80%-90%. Simultaneously, the method disclosed by the invention has the advantages of low taking quantity of the blood, simple operation steps, low cost of a reagent and short maturation time of the cells, above 1*10<9> cells can be achieved by 8 days and used for performing continuous intravenous infusion on a patient six times, and above 1*10<9> cells can be infused every time. The method is suitable for a clinical biological immunotherapy of the patients with tumors and can be widely popularized and applied.

Description

technical field [0001] The invention relates to a method for culturing killer T cells in vitro. More specifically, the present invention relates to a method for inducing, stimulating and culturing human cytotoxic T cells in vitro by using IL-2, anti-CD3 monoclonal antibody and Folium Folium water decoction. Background technique [0002] As the fourth mode of modern cancer treatment, tumor biotherapy based on tumor immunotherapy has been paid more and more attention. Tumor immunotherapy includes active specific immunotherapy (tumor vaccine) and passive immunotherapy, in which passive immunotherapy includes cytokine therapy (interleukin, interferon, tumor necrosis factor, etc.), antibody-based passive immunotherapy, adoptive cell Immunotherapy (such as LAK, TIL, CIK, etc.) and gene therapy. Among them, adoptive cellular immunotherapy (ACI) refers to the transfer of immune cells (specific and relatively specific) with anti-tumor activity to tumor patients to directly kill tum...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783
Inventor 毕薇薇
Owner JILIN TUO HUA BIOTECH
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