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Microfluidic chip for circulating tumor cell separation, circulating tumor cell separation method and counting method

A microfluidic chip and tumor cell technology, which is applied in the direction of tumor/cancer cells, sterilization methods, and biological agents for removing bad cells, can solve problems such as high cost, serious, white blood cell contamination, etc. Improve efficiency and lower cost

Pending Publication Date: 2017-08-29
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

First, there is low or no expression of epithelial adhesion molecules, which leads to missed detection of circulating tumor cells, which fundamentally affects the accuracy of biochemical methods for detecting circulating tumor cells; second, epithelial adhesion molecules and antibody specificity The binding time is long, and the blood sample processing speed is slow; third, the blood sample processing volume is large, more antibodies are required, and the cost is high
Using physical methods to separate circulating tumor cells, the main disadvantages are that the purity of tumor cells is not high, and the contamination of white blood cells is serious, which eventually leads to false positives in the staining process, that is, the occurrence of false detections.
The above defects limit the clinical promotion and application of these technologies

Method used

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  • Microfluidic chip for circulating tumor cell separation, circulating tumor cell separation method and counting method
  • Microfluidic chip for circulating tumor cell separation, circulating tumor cell separation method and counting method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] A method for separating circulating tumor cells using the microfluidic chip for separating circulating tumor cells, the method comprising the following steps:

[0049] (1) Open the input interface of the first shell and the output interface of the second shell, close the output interface of the first shell and the input interface of the second shell, and dilute the peripheral blood of the human body 5 times with phosphate buffer saline Finally, take 1 milliliter of sample, input it from the input interface of the first shell with a pressure of 40mbar, and discharge it from the output interface of the second shell after filtering;

[0050] (2) Open the input interface of the second shell and the output interface of the second shell, close the output interface of the first shell and the input interface of the first shell, and 800 μ L of phosphate buffer solution from the input interface of the second shell input, discharged from the output interface of the second shell; ...

Embodiment 2

[0058] A method for separating circulating tumor cells using a microfluidic chip for the separation of circulating tumor cells, except that the above steps (2) to (5) are not repeated after step (5), and other conditions are the same as those of the implementation Example 1 is the same.

[0059] After 1 filter, the number of particles in the collected sample was 9.63×10 8 , The filtration efficiency of blood cells in the sample by one filtration operation is 86.79%.

Embodiment 3

[0061] A method for counting circulating tumor cells using a microfluidic chip for separating circulating tumor cells, said method comprising the following steps:

[0062] (1') Open the input interface of the first shell layer and the output interface of the second shell layer, close the output interface of the first shell layer and the input interface of the second shell layer, and reduce the concentration to about 1×10 6 After each milliliter of SK-BR-3 cells is diluted 10,000 times with phosphate buffer, 600 μL of sample is input from the input interface of the first shell, filtered and discharged from the output port of the second shell;

[0063] (2') Open the input interface of the second shell and the output interface of the second shell, close the output interface of the first shell and the input interface of the first shell, 100 μ L of phosphate buffer saline is input from the second shell The interface input is discharged by the output interface of the second shell; ...

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Abstract

The invention provides a microfluidic chip for circulating tumor cell separation. The microfluidic chip comprises a first shell layer, a second shell layer and a filter film between the first and second shell layers. A first passage is formed between the filter film and the first shell layer. A second passage is formed between the filter film and the second shell layer. The first shell layer is provided with m input interfaces and n output interfaces, m is greater than or equal to 1 and n is greater than or equal to 1. The second shell layer is provided with x input interfaces and y output interfaces, x is greater than or equal to 1 and y is greater than or equal to 1. The chip has high flux and high efficiency of circulating tumor cells, is used simply and is easy to promote.

Description

technical field [0001] The invention belongs to the technical field of cell separation, and relates to a microfluidic chip for separating circulating tumor cells, and also relates to a method for separating and counting circulating tumor cells. Background technique [0002] Circulating tumor cells are a general term for all kinds of tumor cells that fall off from the primary tumor lesion and enter the peripheral blood of the human body. They are a sign of tumor metastasis. It is of great significance for research directions such as chemical medicine and tumor single-cell sequencing. Related research work has shown that the level of circulating tumor cells in patients is positively correlated with progression-free survival and overall survival. At present, liquid biopsy technology of circulating tumor cells has been clinically applied in the diagnosis of colon cancer, breast cancer, and prostate cancer, and has great application value and market prospect. The content of cir...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/12C12N5/09C12Q1/06
CPCG01N33/5005C12M23/16C12M25/02C12N5/0694B01L2200/0652B01L2300/0681B01L3/502753B01L2200/0668C12M47/04G01N15/1484G01N2015/1006G01N2015/1486G01N15/0625C12N2521/00C12N5/0093G01N15/149B01L3/502761
Inventor 程鑫姜有为余振铭黄兴隆陈日飞
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA