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Morphology and purity and immunophenotyping detection method of human (gamma)(delta)T cells

An immunophenotyping and detection method technology, applied in the field of cellular immunology, to achieve the effects of large quantity, reduced culture cost and high purity

Active Publication Date: 2014-02-12
SHENZHEN HORNETCORN BIOTECH
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  • Claims
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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

[0010] 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.

[0011] Firstly, the present invention uses cultured Mycobacterium tuberculosis to prepare Mycobacterium tuberculosis heat-resistant antigen. Include the following steps:

[0012] 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 morphology and purity and immunophenotyping detection method of human (gamma)(delta)T cells, which is used for detecting the prepared human (gamma)(delta)T cells. The method comprises the following steps: preparing human (gamma)(delta)T cells; and performing morphological observation with a microscope and purity and immunophenotyping analysis with a flow cytometer. The method provided by the invention has the advantages that the (gamma)(delta)T cells obtained by the test have the characteristics of large quantity, high purity, strong cytotoxicity and the like so as to ensure that the prepared human (gamma)(delta)T cells can meet the clinical needs; and moreover, the effects that the culture cost is greatly reduced from the non-peptide phosphate antigens such as IPP and the irritants such as anti-TCR (gamma)(delta) antibody are realized and the problems of small quantity, low toxicity, high cost and the like of in-vitro culture of (gamma)(delta)T cells in the prior art are solved.

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 morphology, purity and immunophenotype of 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% o...

Claims

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Application Information

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IPC IPC(8): C12N5/0783G01N33/569
Inventor 马飞王宇环罗晓玲
Owner SHENZHEN HORNETCORN BIOTECH
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