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Preparation method of dendritic nanotube array, method for identifying tumor cells, and microfluidic device for capturing and in situ regulating cancer cells

A nanotube array, tumor cell technology, applied in tumor/cancer cell, tissue cell/virus culture devices, biochemical equipment and methods, etc., can solve problems such as cognition of the mechanism that restricts the activity of cancer cells, and achieve excellent biosafety Sexuality, the effect of precise drug regulation

Active Publication Date: 2019-11-29
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] At present, most of the CTCs microfluidic chips are more limited to capturing, separating and releasing cancer cells, and few of them perform more complex and more precise and controllable operations on the captured cancer cells in situ, such as releasing drugs and regulating cancer cells. cell behavior
Limitations of these cancer cell in situ regulation and detection assays limit understanding of the mechanisms of cancer cell activity
In the research and development of CTCs detection technology, there is still a lack of a method that can capture and separate cancer cells, as well as in situ drug regulation of cancer cell behavior

Method used

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  • Preparation method of dendritic nanotube array, method for identifying tumor cells, and microfluidic device for capturing and in situ regulating cancer cells
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preparation example Construction

[0040] The preparation method of a kind of branched nanotube array of the present invention: its specific steps are:

[0041] (1) Using atomic layer deposition (ALD for short) technology, gas-phase diethyl zinc is used as a precursor and mixed with water vapor into the reaction chamber, and a ZnO film is deposited on the surface of the substrate film and the inner wall of the hole. The thickness value of the ZnO thin film is 30~40nm;

[0042] (2) continue to feed trimethylaluminum and water vapor in the gas phase for a period of time, and deposit Al on the surface of the ZnO film 2 o 3 nanolayer, the Al 2 o 3 The thickness of the nano-layer is 10-20nm;

[0043] (3) in BCl 3 and Cl 2 Under the mixed gas, the plasma etched away the Al on the surface of the substrate film 2 o 3 / ZnO layer, followed by O 2 Part of the polycarbonate film is etched away by plasma to expose the nanotube array structure. At this time, by changing the O 2 The length of the plasma etching time...

Embodiment 1

[0063] Human breast cancer cells (MCF7) were selected as a model system for cell capture, and 50 μL containing 10 6 The cells / mL MCF7 cell suspension was injected into the upper cell culture tank 1 of the microfluidic device, placed in the cell culture incubator and incubated for 60 minutes, then washed with a large amount of cell culture medium, and observed under a fluorescence microscope, a large number of cells can be found Captured, the capture efficiency can reach more than 90%.

Embodiment 2

[0065] To study the specific detection ability of dendritic nanotubes:

[0066] Human breast cancer cells (MCF7) and human prostate cancer cell lines (PC-3) were used as EpCAM positive cell line controls, and human cervical cancer cell lines (Hela) and human T lymphocytes (Jurkat T) were used as EpCAM negative cell line controls. 50 μL containing 10 6 The cells / mL cell suspension was injected into the upper cell culture pool 1 of the microfluidic device, placed in the cell culture incubator and incubated for 90 minutes, and then washed with a large amount of cell culture medium, observed under a fluorescence microscope, it can be found that EpCAM is positive In the cell line group, a large number of cells were captured, and the capture efficiency was more than 90%; in the EpCAM-negative cell line group, a small number of cells were captured, and the capture efficiency was less than 40%.

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Abstract

The invention belongs to the technical field of medical detection of cancer cells, and particularly relates to a preparation method of a nanotube array, a method for specially identifying circulatingtumor cells through a dendritic nanotube array and a microfluidic device used for capturing and regulating cancer cells in situ through the nanotube array. The microfluidic device comprises a PDMS module pool, wherein the PDMS module pool comprises a cell culture pool and a microfluidic channel, and substrate film of the dendritic nanotube array is used for partitioning the cell culture pool on the upper layer and the microfluidic channel on the bottom layer; and the two tail ends of the microfluidic channel are provided with input / output ports used for conveying solutions. The preparation method of the dendritic nanotube array, the method for specially identifying the circulating tumor cells through the dendritic nanotube array and the microfluidic device used for capturing and regulatingthe cancer cells in situ through the nanotube array have the advantages that the capture of the cancer cells has high efficiency and strong specificity, antibody molecules identified specially are modified on a dendritic nanotube, the cancer cells are effectively captured and separated from blood, the dendritic nanotube has excellent biological safety for the cells, and the normal function of thecancer cells is not affected.

Description

technical field [0001] The invention belongs to the technical field of detection of medical cancer cells, in particular to a method for preparing a dendritic nanotube array, a method for specifically identifying circulating tumor cells by using a dendritic nanotube array, and using the nanotube array to capture and regulate cancer cells in situ. Microfluidic devices for cells. Background technique [0002] In the prior art, cancer is one of the major diseases threatening human health and life in the world. Biomedical research has been committed to revealing the mechanism of cell carcinogenesis, developing cancer diagnosis technology, and proposing cancer prevention and treatment programs, developing new cancer cell detection technology, and realizing drug regulation of cancer cell behavior, which is crucial to the development of new precision and personalized medicine. Cancer treatment methods are of great value. [0003] Traditional tumor diagnosis methods mainly include ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N33/543G01N33/574C12N5/09C12M3/00C12M1/00
CPCC12M23/16C12N5/0693G01N33/54346G01N33/57488
Inventor 谢曦何根冯键铭周灵菲文瑞黄爽杨成端杭天陈惠琄刘繁茂杨柏儒
Owner SUN YAT SEN UNIV
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