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New application of near-infrared II area quantum dots containing selenium elements

A quantum dot and near-infrared technology, which is applied in the preparation of in vivo experiments, blood/immune system cells, vertebrate cells, etc., can solve the long-term tracking of unfavorable cell migration, increase the cost of probe preparation, and the price of penetrating peptides Expensive and other issues, to achieve the effect of simple and convenient marking process, stable marking effect and good marking effect

Active Publication Date: 2018-09-28
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

For example, CD3 antibody can be modified on the surface of quantum dots, and T cell labeling can be completed by co-culturing with T cells for 30 minutes to obtain a better labeling effect. The disadvantage is that the antibody is expensive, the material needs to be modified, and the operation is cumbersome. The reaction with the ligand occurs only on the cell surface and the process is reversible, resulting in unstable labeling and possible shedding during tracking
Quantum dots modified by membrane-penetrating peptides can efficiently enter the interior of cells, and the labeling effect is stable. However, membrane-penetrating peptides are expensive, which greatly increases the cost of probe preparation, and the mechanism of action of membrane-penetrating peptides is to forcibly penetrate the membrane. Has a damaging effect, which is not conducive to long-term tracking of cell migration

Method used

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  • New application of near-infrared II area quantum dots containing selenium elements
  • New application of near-infrared II area quantum dots containing selenium elements
  • New application of near-infrared II area quantum dots containing selenium elements

Examples

Experimental program
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Effect test

Embodiment 1

[0056] (1) Extraction and culture of mouse T cells

[0057] Take the spleen of 4-6 weeks old Balb / c mouse, cut it into pieces and grind it on a 200-mesh cell mesh sieve to obtain a spleen homogenate, dilute the spleen homogenate to 5mL and transfer it to a 15mL centrifuge tube, carefully add an equal volume of spleen lymph Cell separation solution, centrifuged at 300g for 30 minutes, sucked out the lymphocyte layer, resuspended with cleaning solution, washed by centrifugation for 3 times, and then purified with nylon cotton column to obtain mouse T cells;

[0058] Inoculate the obtained T cells into culture plates or cell culture flasks, culture them in 1640 medium (containing 10% fetal bovine serum, 0.5% double antibody, 25mM HEPES buffer and 2mM L-glutathione), and add 50ng / mL of CD3 antibody and 200IU / mL interleukin-2 were cultured in a cell incubator, and the medium was replaced every two days, and interleukin-2 was added.

[0059] (2) Marking of T cells

[0060] Centri...

Embodiment 2

[0068] Compared with Example 1, T cells were labeled with PbSe near-infrared II region quantum dots, and other conditions were the same as Example 1. The experimental results except that due to PbSe and Ag 2 Except for the difference in fluorescence signal intensity caused by the different quantum yields of the two quantum dots of Se, there is no qualitative difference from Example 1.

[0069] In summary, the present invention uses the natural selenium binding transport system of T cells to allow T cells to take in the near-infrared II region quantum dots containing selenium, and realizes the natural, non-destructive and highly efficient treatment of T cells by the near-infrared II region quantum dots. Quantum dots in the near-infrared II region are evenly distributed in T cells, the labeling effect is stable, and will not be interfered by biological autofluorescence in in vivo fluorescence imaging, with strong tissue penetration ability and high spatial resolution; the presen...

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Abstract

The invention provides new application of near-infrared II area quantum dots containing selenium elements. According to the new application, the near-infrared II area quantum dots are used for T cellmarking and / or T cell in-vivo tracking and contain the selenium elements. According to the new application of the near-infrared II area quantum dots containing the selenium elements, by utilizing natural selenium element transfer systems of T cells, the T cells take in the near-infrared II area quantum dots containing the selenium elements, natural, nondestructive and efficient marking of the T cells is achieved, and the marked quantum dots are evenly distributed in T cell cytoplasm; by utilizing a laser confocal microscope or a small animal in-vivo imaging system, in-vivo tracking of the T cells is achieved through near-infrared II area fluorescence signals; and efficient and nondestructive marking of the T cells can be achieved without using expensive bioactive molecules to modify the quantum dots.

Description

technical field [0001] The invention belongs to the technical field of biological imaging and relates to a new application of quantum dots in the near-infrared II region containing selenium element. Background technique [0002] T cells are a type of lymphocytes and are an important component of the adaptive immune system. With the assistance of antigen-presenting cells, T cells can recognize and kill virus-infected cells or cells with abnormal gene expression (such as cancer cells), and play an important role in fighting against external pathogen invasion and tumor immunotherapy. [0003] Tumor immunotherapy is an emerging precise and effective tumor treatment method, mainly including immune node check inhibitor therapy and adoptive T cell therapy. Among them, adoptive T cell therapy is to modify T cells from the patient's own or allogeneic body in vitro to make them have the ability to kill tumors, and then infuse them back into the patient's body to achieve the killing e...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0783A61K49/00
CPCA61K49/0067A61K49/0097C12N5/0636
Inventor 王强斌陶惠泉
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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