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Cyan fluorescent protein marked tumor cell strain, and construction method and application thereof

A cyan fluorescent protein and tumor cell line technology, which is applied in the field of cyan fluorescent protein-labeled tumor cell line construction, can solve the problem of restricting the use of tumor cells and transgenic mice

Inactive Publication Date: 2017-12-19
HUAZHONG UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Currently, the common transgenic mice with fluorescent protein-labeled immune cells, such as CX3CR1-GFP, CXCR6-EGFP, and Foxp3-mRFP, mostly express GFP and RFP, which limits the relationship between fluorescent protein-labeled tumor cells and transgenic mice. When used together, there are certain limitations in the process of realizing multi-color observation of the tumor immune microenvironment

Method used

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  • Cyan fluorescent protein marked tumor cell strain, and construction method and application thereof
  • Cyan fluorescent protein marked tumor cell strain, and construction method and application thereof
  • Cyan fluorescent protein marked tumor cell strain, and construction method and application thereof

Examples

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Embodiment 1

[0023] Example 1: Tumor cell lines labeled with cyan fluorescent protein

[0024] An embodiment of the present invention provides a cyan fluorescent protein-labeled tumor cell line: a tumor cell line is constructed by stably transfecting a cyan fluorescent protein particle into a tumor cell line. Among them, cyan fluorescent protein, the English full name is cyanfluorescentprotein, abbreviated as CFP.

Embodiment 2

[0025] Example 2: Construction method of cyan fluorescent protein-labeled tumor cell line

[0026] see figure 1 As shown, the embodiment of the present invention also discloses a method for constructing a cyan fluorescent protein-labeled tumor cell line, comprising the following steps:

[0027] S1, using pcDNA3.1 as a plasmid vector to prepare a plasmid expressing cyan fluorescent protein, see figure 2 Shown; Tumor cell lines were transfected with plasmids expressing cyan fluorescent protein;

[0028] S2, the cells were cultured and screened in G418 complete medium to obtain a cyan fluorescent protein-labeled tumor cell line.

Embodiment 3

[0029] Example 3: Application of Cyan Fluorescent Protein-labeled Tumor Cell Lines

[0030] The embodiment of the present invention also discloses the application of a cyan fluorescent protein-labeled tumor cell line, the tumor cell line is a B16 tumor cell line, and the cyan fluorescent protein-labeled tumor cell line is used for co-incubation with specific CTL in vitro to Observe the whole process of killing tumor cells by specific CTL.

[0031] Cyan fluorescent protein-labeled tumor cell lines are used to inject into the animal skin window of various fluorescent protein-labeled immune cells, and combined with DiR-BOA fluorescent dye to label blood vessels to dynamically observe tumor cells and immune cells in the tumor microenvironment in vivo.

[0032] Cyan fluorescent protein-labeled tumor cell lines are used for injection into animal skin windows to observe the tumor treatment effect after adopting tumor treatment regimens.

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Abstract

The invention discloses a cyan fluorescent protein marked tumor cell strain, and a construction method and the application thereof, and relates to the technical fields of tumor immunology and optical imaging. The cyan fluorescent protein marked tumor cell strain is obtained through cyan fluorescent protein plasmid transfection and G418 medicine screening, so that the application of the cyan fluorescent protein marked tumor cell strain to tumor immunity optical research is realized, for example, the whole process of killing tumor cells by CTL is observed and researched through microscopic optical imaging, or multicolor marking and microscopic dynamic imaging of the tumor microenvironment are realized in vivo, or real-time dynamic optical imaging observation on the dependent interaction among the tumor cells, various immune cells and blood vessels in the tumor microenvironment is realized in the tumor treatment process.

Description

technical field [0001] The invention relates to the technical fields of tumor immunology and optical imaging, in particular to a cyan fluorescent protein-labeled tumor cell line, a construction method and an application. Background technique [0002] In recent years, tumor immunotherapy has become the most popular field in cancer treatment research, and is recognized as the most likely treatment method to overcome cancer. What cellular and molecular events occur at what time and where in the process of tumor immunity is still poorly understood, and the ever-developing optical imaging technology has the characteristics of high sensitivity, high temporal and spatial resolution, multi-parameter detection and low damage ( See Luo Qingming, Zhang Zhihong. Acta Optics Sinica, 2011,31(9):119-125.), it has played a huge potential in the study of tumor immunology, it can provide information on the interaction process between tumor cells and immune cells in the process of tumor immuni...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10C12N15/65A61K49/00G01N21/64
CPCC12N5/0693A61K49/0045C07K14/43595C12N15/65G01N21/6486
Inventor 张智红祁淑红
Owner HUAZHONG UNIV OF SCI & TECH
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