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Equipment, system and method for separating CTCs (circulating tumor cells)

A technology for tumor cells and separation equipment, applied in the direction of tumor/cancer cells, biochemical equipment and methods, animal cells, etc., can solve the problems of low purity of CTC, prone to false positives, and inability to release detection, etc., to improve the purity of CTC, The effect of maintaining CTC activity and improving purity

Inactive Publication Date: 2017-10-24
WUXI NAAO BIO PHARM
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1) The purity of captured CTCs is low, there is non-specific adsorption, and false positives are prone to occur
[0006] 2) The captured tumor cells are immobilized on the surface of the chip and cannot be released for subsequent detection

Method used

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  • Equipment, system and method for separating CTCs (circulating tumor cells)
  • Equipment, system and method for separating CTCs (circulating tumor cells)
  • Equipment, system and method for separating CTCs (circulating tumor cells)

Examples

Experimental program
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Effect test

Embodiment 1

[0030] Such as figure 1 As shown, the circulating tumor cell capture device provided by the present invention includes a control module (1: control screen, 7: control board), a sampling module (2: a sampling needle and a stirring rod), a separation module (5: a magnetizer, 3 : separator), collection module (4: sample collection rack).

[0031] The separator is composed of a plurality of single-channel microfluidic chips. The microfluidic chip is formed by reversible sealing of the upper PDMS polymer chip and the lower silicon chip carrier. It is a spiral channel with a channel width of 600 microns. The height is 100 microns and the length is 50 cm. The inner wall of the microfluidic chip is coated with amphiphilic polymer PEG-PCL. Sample port (circulating tumor cell sample port and waste liquid sample port), the cell sample after removing white blood cells enters the microfluidic chip from the sample port, and the separated circulating cell suspension is transported from the...

Embodiment 2

[0041] Five patients diagnosed with pancreatic cancer were selected. After the ethical approval and informed consent of the patients, 7.5 ml of peripheral blood was drawn from each and placed into EDTA anticoagulant tubes.

[0042] Circulating tumor cell isolation procedure:

[0043] 1) Sample preparation: after red blood cell lysis, centrifuge at 1700rpm, remove the lysed red blood cells, and redisperse in a 50ml centrifuge tube (sample tube) with 5ml of PBS containing 0.5%BSA and 2mM EDTA (pH7.5) , while adding 50ul1mg / ml immunomagnetic particles.

[0044] 2) Put the sample tube into the sample rack of the equipment Nextctc, the instrument starts to run the program for 1 minute, the control screen prompts to enter the patient information, after setting the operating parameters, click start. The instrument injects the diluent, the stirring motor runs, and stabilizes for 15 minutes. The magnet pushes the motor to run to adsorb the magnetic beads to the wall of the sample tube...

Embodiment 3

[0048] Select 5 patients with breast cancer diagnosis, after the ethical approval and the informed consent of the patients, 5ml of peripheral blood was drawn from each of them, and put into EDTA anticoagulant tubes.

[0049] Circulating tumor cell isolation procedure:

[0050] 1) Sample preparation: after red blood cell lysis, centrifuge at 1800rpm, remove the lysed red blood cells, and redisperse them in a 50ml centrifuge tube (sample tube) with 5ml of PBS containing 0.5%BSA and 2mM EDTA (pH7.5). , while adding 80ul1mg / ml immunomagnetic particles.

[0051] 2) Put the sample tube into the sample rack of the equipment Nextctc, the instrument starts to run the program for 1 minute, the control screen prompts to enter the patient information, after setting the operating parameters, click start. The instrument injects the diluent, the stirring motor runs, and stabilizes for 15 minutes. The magnet pushes the motor to run to adsorb the magnetic beads to the wall of the sample tube....

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Abstract

The invention belongs to the field of biological and medical detection and relates to equipment, a system and a method for separating CTCs (circulating tumor cells) in a blood sample from mixed cell groups with high purity. The equipment and system for separating the CTCs comprise, but not limited, a control module, a sample injection module, a separation module and a collection module, wherein the control module is used for setting equipment operation parameters including diluents volume, reaction time, sample injection speed, cleaning parameter and the like; the sample injection module adopts a multi-way valve module and a peristaltic pump module and is used for automatically diluting detected samples and starting multi-channel synchronous separation of the samples; the separation module comprises a magnet module and a separator module, the magnet module is used for separating and adsorbing magnetic particle immunolabeled blood leukocytes, and the separator module is used for enriching and separating CTCs from multiple cell groups; the collection module is used for collecting a separated CTC suspension. The invention provides the equipment and the system for separating the CTCs as well as the method for separating and identifying the CTCs.

Description

technical field [0001] The invention relates to a circulating tumor cell separation device, system and method. Background technique [0002] Circulating tumor cells (CTC) are circulating cancer cells that come off from the original cancer cell tissue and enter the peripheral blood and tissues of the human body. CTC is of great significance for the early diagnosis of cancer and the evaluation of treatment effect. Due to the extremely rare number of CTCs, each 10mL of blood may only contain a few to dozens of circulating tumor cells, but there are as many as 100 million white blood cells and 50 billion red blood cells (ZheXN, CherML, BonfilRD, Am.J.Cancer Res.2011 , 1, 740-751), under the interference of a huge number of background cells, it is impossible to directly detect circulating tumor cells. This detection includes gene mutation, surface markers, cell activity, secreted protein profile and mechanical properties, etc., so CTC Isolation from peripheral blood becomes a ne...

Claims

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Application Information

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IPC IPC(8): C12M1/36C12M1/02C12M1/00C12N5/09
CPCC12M23/16C12M23/20C12M41/48C12M47/04C12N5/0693
Inventor 亓立峰
Owner WUXI NAAO BIO PHARM
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