An automatic separation and enrichment device and method for tumor cells

A technology for separating and enriching tumor cells, which is applied in the field of automated separation and enrichment devices, and can solve the problems of clinical diagnosis deviation, low efficiency of separating whole blood, and cell damage.

Active Publication Date: 2021-09-10
睿思生命(广东)科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] 1. The separation and enrichment process is complicated, and the operator's participation is high, which increases the probability of errors in the intermediate links, thus requiring higher professional skills for personnel;
[0006] 2. The recovery rate of the target cells is low, resulting in false negative results and deviations in clinical diagnosis;
[0007] 3. The efficiency of separating whole blood is low, the sample separation time is long, and the cells undergo more pretreatment, which largely causes cell damage and loss of activity

Method used

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  • An automatic separation and enrichment device and method for tumor cells
  • An automatic separation and enrichment device and method for tumor cells
  • An automatic separation and enrichment device and method for tumor cells

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Experimental program
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Embodiment 1

[0060] An operation method of an automated microfluidic system for enrichment of circulating tumor cells, the method adopts the method attached to the present invention Figure 1-4 equipment, the specific operations are as follows:

[0061] S01: Remove the sample tube and the buffer tube in the device respectively, add 2.3mL of ten-fold concentration of cell preservation solution to the buffer tube, then add deionized water to the buffer solution to the set scale, and mix well;

[0062] S02: Put the sample or blood collection tube to be tested into the sample tube, and add the mixed cell preservation solution to the sample tube to 5mL;

[0063] S03: Put the buffer tube and sample tube into the corresponding position of the device, put a 50mL centrifuge tube in the waste tube rack as the waste tube, and then put a 1.5mL or 2mL enrichment tube in the enrichment tube slot;

[0064] S04: Press the start button, wait for the sample to be loaded, and after the sample in the sample ...

Embodiment 2

[0067] Adopt the present invention to attach Figure 1-4 The equipment in the method separates different types of cell lines, and recovers the separated cell lines and calculates the recovery efficiency of the cell lines. The specific method is as follows:

[0068] 1) Take the non-small cell lung cancer cell line H1299 cultured in T25, discard the medium, add 5mL PBS to wash twice, add 1mL trypsin to make the trypsin evenly cover the cell surface, discard the excess trypsin, and incubate at 37 degrees Celsius for 3 Minutes, add medium containing 10% FBS to terminate the digestion reaction, mix the cells evenly with a 1mL pipette, and confirm under the microscope that all cells are dispersed into single cells. And take 100 μL of cells, use a cell counter for counting.

[0069] 2) According to the concentration calculated by the cell counter, use the cell preservation solution to dilute H1299. The dilution concentration is 100 H1299 cells in the 5mL dilution solution. Add the dil...

Embodiment 3

[0077] Adopt the present invention to attach Figure 1-4 The device in the method separates the cells and detects the activity of the separated cells, the specific method is as follows:

[0078] 1) Take the lung cancer cell line A549 cultured in T25, discard the medium, add 5mL PBS to wash twice, add 1mL trypsin to make the trypsin evenly cover the cell surface, discard the excess trypsin, incubate at 37 degrees Celsius for 3 minutes, add The medium containing 10% FBS stopped the digestion reaction, mixed the cells evenly with a 1mL pipette, and confirmed under the microscope that all the cells were dispersed into single cells. And take 100 μL of cells, add trypan blue according to 1:1, use a cell counter to count and calculate the cell viability; the measured initial cell viability is 96.81%.

[0079] 2) Adjust the cell concentration to 1×106 cells / mL, and add the diluted cells to the attached Figure 1-3 in the sample tube of the device and use the device for separation. ...

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Abstract

The invention discloses an automatic separation and enrichment device and method for tumor cells. In the device, the inlet end of the chip is connected with a buffer unit and a sample unit, the outlet end of the chip is connected with a enrichment liquid unit and a waste liquid unit, and the power unit is simultaneously Connected to the sample unit and the buffer unit; the chip is a microfluidic chip; when the buffer solenoid valve, the sample pinch valve and the buffer pinch valve are opened, the pressure generated by the power unit forces the buffer in the buffer tube into the sample tube; When the sample solenoid valve, sample pinch valve, buffer solenoid valve, buffer pinch valve, enrichment pinch valve, and waste liquid pinch valve are open, the pressure generated by the power pack forces the sample in the sample tube and the buffer tube in the buffer tube The buffer solution passes through the chip into the enrichment tube and the waste tube respectively. The invention can reduce damage to cells, recover more target cells, ensure the reliability of circulating tumor cells in downstream analysis; and shorten separation time and improve detection efficiency.

Description

technical field [0001] The invention relates to the field of cell sorting and enrichment, in particular to an automatic separation and enrichment device and method for tumor cells. Background technique [0002] Circulating tumor cells (CTCs) are tumor cells that enter the peripheral blood from the primary tumor lesion or metastatic lesion. Through accurate detection of CTCs, it can be effectively used for early diagnosis in vitro, rapid evaluation of chemotherapy drugs, and helpful for better treatment of patients. Facilitate individualized treatment. In particular, the process of obtaining non-destructive living cells of CTCs is of great significance. The non-destructive living cell culture of CTCs can provide valuable replica samples for the study of CTCs. However, due to the scarcity of CTCs in the peripheral blood of tumor patients, studies have shown that there is only 1 CTCs in 10^6-10^7 monocytes on average. In addition to the complexity of the blood, how to separate...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12M1/36C12M1/34C12M1/04C12N5/09B01L3/00
CPCB01L3/5027C12M23/16C12M29/06C12M41/40C12N5/0693
Inventor 崔彩媚张雷丁汝波甘剑亮
Owner 睿思生命(广东)科技有限公司
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