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Method for promoting targeting conveying effect of nanometer substance in living cells

A live cell, targeted technology, applied in the field of biomedicine, which can solve the problems of poor efficiency and high cytotoxicity

Inactive Publication Date: 2019-03-01
NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Existing methods used to help nanomaterials escape from vesicles are often inefficient or toxic to cells, or both

Method used

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  • Method for promoting targeting conveying effect of nanometer substance in living cells
  • Method for promoting targeting conveying effect of nanometer substance in living cells
  • Method for promoting targeting conveying effect of nanometer substance in living cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 The method and effect of DMSO on TAT-QD increasing target in cervical cancer tumor cells.

[0043] The specific operation steps are as follows: after the HeLa cells adhere to the wall, normal cell culture medium (DMEM medium containing 10% fetal bovine serum) and cell culture medium containing 1% DMSO in volume concentration are respectively selected. The TAT-QDs were dispersed in the above two culture solutions at the same concentration. After incubation with the cells in an incubator (37°C, 5% CO 2 ) after co-cultivation for the indicated time, observed under confocal laser microscope.

[0044] In order to colocalize the relative positions of organelles and quantum dots, we stained the nucleus with the nuclear dye Hoechst 33342, and stained the cell membrane and inclusion bodies with DiO. Cell nuclei staining method: Remove the cell culture medium and wash once with PBS. Add 600 μL of Hoechst 33342 staining solution (dissolved in DMEM) at a concentration ...

Embodiment 2

[0047] Embodiment 2 A variety of cosolvents can improve the targeting effect of TAT-QD

[0048] The specific operation is: after the HeLa cells adhere to the wall, select normal cell culture medium (DMEM medium containing 10% fetal bovine serum) and cells containing a cosolvent (DMF, ethanol, acetone or tetrahydrofuran) with a volume concentration of 1%. culture medium. The TAT-QDs were dispersed in the above two culture solutions at the same concentration. After incubation with the cells in an incubator (37°C, 5% CO 2 ) after co-cultivation for 24 hours, observed under confocal laser microscope. In order to co-localize the relative positions of organelles and quantum dots, we stained the nuclei with the nuclear dye Hoechst 33342.

[0049] Such as Figure 4 It shows that, like DMSO, TAT-QD can escape from the vesicles and enter the nucleus with the participation of DMF, ethanol, acetone or THF, but the effect of THF is slightly worse than that of other co-solvents. . It ...

Embodiment 3

[0050] Example 3 The targeting effect of TAT-QD improved by cosolvent can be realized in various cell types

[0051] The specific operation steps are: select human breast cancer cells (MCF-7), human liver cancer cells (Hep G2) and human lung cancer cells (A549), and after the cells adhere to the wall, select normal cell culture medium (containing 10% fetal bovine Serum culture medium) and cell culture medium containing 1% DMSO in volume concentration. The TAT-QDs were dispersed in the above two culture solutions at the same concentration. After incubation with the cells in an incubator (37°C, 5% CO 2 ) after co-cultivation for the indicated time, observed under confocal laser microscope. In order to co-localize the relative positions of organelles and quantum dots, we stained the nuclei with the nuclear dye Hoechst 33342.

[0052] Such as Figure 5 It was shown that, like HeLa cells, TAT-QD could escape from the vesicles and enter the nucleus with the participation of coso...

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Abstract

The invention relates to a method for promoting a targeting conveying effect of a nanometer substance in living cells and belongs to the technical field of biomedicine. The method comprises the following steps: 1) preparing nano-particles with targeting effect; 2) taking the nano-particles with targeting effect by cells; 3) observing distribution of quantum dots in cells in the manner of fluorescent co-localization; 4) adopting a MTT method for determining cytotoxicity. According to the method for promoting the targeting conveying effect of the nanometer substance in living cells provided by the invention, the nanometer substance can be effectively helped to enter into cells and then escape from vesicae and enter into targeting organelle positions. The toxicity to cells can be almost ignored.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a method for improving the targeted delivery effect of nano-level substances in living cells. Background technique [0002] In the field of biomedicine, nanomaterials (such as carbon nanotubes, silicon nanowires, gold nanoparticles, quantum dots, etc.) have been widely used; in addition, various biological macromolecules (proteins, SiRNA, etc.) in the field of biomedicine. [0003] The targeted labeling technology of these nanoscale substances on the cell surface is relatively easy, but the targeted delivery inside living cells is a difficult problem. This is because, in order to achieve the purpose of targeted delivery in living cells (such as delivery to a specific organelle), nanomaterials need to cross multiple transport barriers. [0004] When the nanomaterial is taken up by the cell through the usual endocytic pathway, it will be trapped in the vesicle and...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K49/00A61K47/62
CPCA61K49/0017A61K49/0056
Inventor 阮刚雍雪青杨轩
Owner NANJING UNIV
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