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Method for detecting killing activity of human (gamma)(delta)T cells against K562 cell strain

A technology of killing activity and detection method, applied in the field of cellular immunology, to achieve the effects of reduced culture cost, large quantity and strong cytotoxicity

Inactive Publication Date: 2014-02-12
SHENZHEN HORNETCORN BIOTECH
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  • Description
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AI Technical Summary

Problems solved by technology

For the detection of human γδT cells

Method used

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Embodiment Construction

[0009] The present invention is specifically described below with examples, but the present invention is not limited thereto. In the following examples, all the experimental methods that do not indicate the specific conditions are implemented in accordance with the conventional methods and the operating instructions provided by the manufacturer.

[0010] Firstly, the present invention uses cultured Mycobacterium tuberculosis to prepare Mycobacterium tuberculosis heat-resistant antigen. Include the following steps:

[0011] 1. Inoculate Mycobacterium tuberculosis seeds with a wet weight of 50-100 grams into 200ml Sutong-type liquid medium, culture them statically in a 37°C incubator for 2 weeks, and then divide them into 10%-30% newborn In 250ml Sutong type liquid medium of bovine serum, 50ml per bottle, divided into 4 bottles, and then continue to culture in a 37°C incubator for 4 weeks, collect the cultured bacterial suspension, centrifuge at 4000 rpm 30 minutes to harvest ...

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Abstract

The invention belongs to the technical field of cellular immunology, and particularly relates to a method for detecting the killing activity of human (gamma)(delta)T cells against a K562 cell strain, which is used for detecting the prepared human (gamma)(delta)T cells. The method comprises the following steps: preparing the human (gamma)(delta)T cells; fetching the K562 cell strains of the logarithmic phase as target cells, and adjusting the cell density to 1*10<5> / ml, 5*10<4> / ml and 2.5*10<4> / ml; paving the target cells in a 96-hole culture plate with 100mu l per hole, and culturing for 24 hours; fetching the prepared human (gamma)(delta)T cells, and adjusting the cell density to 1*10<6>ml; adding the human (gamma)(delta)T cells into the 96-hole culture plate, wherein the effect-target ratios are 10:1, 20:1 and 40:1 respectively; and setting 3 parallel holes for each density, setting blank controls respectively and calculating the killing activity. By adopting the method provided by the invention, the toxicity of the human (gamma)(delta)T cells is guaranteed.

Description

[0001] This application is a divisional application of the invention patent "A Simple and Efficient Method for Preparing Human γδT Cells" with application number 201210030197.4 dated February 10, 2012. technical field [0002] The invention belongs to the technical field of cellular immunology, and specifically relates to a method for detecting the killing activity of K562 cell strain by a large number of predominantly amplified human peripheral blood γδT cells in vitro. Background technique [0003] T lymphocytes are divided into two types according to the expression of T cell antigen receptor (TCR): TCRαβT cells and TCRγδT cells, referred to as αβT cells and γδT cells respectively. Among them, γδT cells are a special type of cells between specific immunity and nonspecific immunity. Most of them are double-negative for CD4 and CD8 molecules, and a few can express CD8 molecules. γδT cells are mainly distributed in skin and mucosal tissues, and generally do not exceed 5% of t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/20C12R1/32
Inventor 马飞王宇环罗晓玲
Owner SHENZHEN HORNETCORN BIOTECH
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