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Identification method of EB (Epstein-Barr) virus transformation cell apoptosis of excellent ice athletes

A technology for Epstein-Barr virus and transformed cells, which is applied in measuring devices, individual particle analysis, scientific instruments, etc., can solve the problems of expensive detection, strong subjectivity, and non-quantitative detection

Inactive Publication Date: 2013-07-10
HARBIN INST OF PHYSICAL EDUCATION
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Problems solved by technology

Apoptosis is usually observed by various staining of tissues or cells by light microscope, fluorescence microscope and electron microscope, followed by in situ terminal labeling (Terminal deoxynucleotidyl trasnferase-mediated deoxyuridine triphosphate nick-end labeling assay, TUNEL) and DNA agarose gel electrophoresis is used to analyze apoptosis, but these methods have the disadvantages of expensive detection, too much subjectivity, and cannot be quantitatively detected.

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  • Identification method of EB (Epstein-Barr) virus transformation cell apoptosis of excellent ice athletes

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Embodiment Construction

[0009] (1) Prepare a single-cell suspension, collect EB virus-transformed cells in a flow tube, and centrifuge at 1200 rpm for 8 minutes.

[0010] (2) Wash the cells twice with phosphate buffered solution (PBS) (centrifuge at 1200rpm for 8min) and collect (1-5)×10 5 cell.

[0011] (3) Add 100 μl of binding buffer (Binding Buffer) to each sample to suspend the cells.

[0012] (4) Add 5 μl FITC and 5 μl PI for double staining at the same time, and mix well.

[0013] (5) Apoptosis was induced by a drug known to induce apoptosis of Epstein-Barr virus-transformed cells (such as camptothecin), as a positive control. Stain with 5 μl FITC and PI respectively before detection.

[0014] (6) At room temperature, protected from light, react for 5-10 minutes.

[0015] (7) First observe the apoptosis of cells under confocal 488nm.

[0016] (8) Add 400 μl Binding Buffer, pass through a 400-mesh sieve, and perform flow cytometry quantitative detection within 1 hour.

[0017] (9) Judgmen...

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Abstract

The invention discloses an identification method of EB (Epstein-Barr) virus transformation cell apoptosis of excellent ice athletes. The implementation steps of the identification method of the EB virus transformation cell apoptosis of the excellent ice athletes are shown in a specific implement mode part. The identification method provided by the invention provides a researching mode for the EB virus transformation cell apoptosis of the excellent ice athletes, the EB virus transformation cell apoptosis conditions of the excellent ice athletes are subjected to quantitative analysis, according to the identification method, viable apoptotic cells, non-viable apoptotic cells and non-viable non-apoptotic cells can be distinguished quantitatively, and the effect and the accuracy are reliable through the electron microscope inspection and Tunel method inspection.

Description

technical field [0001] The invention relates to a method for identifying the apoptosis of excellent ice and snow athlete Epstein-Barr virus (epstein-barr virus, EB) transformed cells by using flow cytometry. Background technique [0002] The technology of cell immortalization has made considerable progress. Epstein-Barr virus and Simian vacuolating virus 40 (SV40) are commonly used transforming viruses for immortalized cells. Epstein-Barr virus transforms the peripheral blood B lymphocytes of outstanding ice and snow athletes to establish an immortal cell line, which allows human cells with a certain lifespan to reproduce uninterruptedly, strengthens the proliferation ability of some special cells, and reduces the apoptosis rate. Epstein-Barr virus-transformed cells of excellent ice and snow athletes can permanently preserve human genetic resources related to outstanding sports ability, and at the same time solve the contradiction between the relay-style conduct of previous ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N15/14
Inventor 陈伟光田小健马喜强王会孙博关伟军
Owner HARBIN INST OF PHYSICAL EDUCATION
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