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Circulation tumor cell separation and enrichment micro-fluidic chip and enrichment method thereof

A microfluidic chip and tumor cell technology, applied in the field of microfluidic chip for separation and enrichment of circulating tumor cells and its enrichment, can solve the problems of undiscovered surface markers of circulating tumor cells, false negatives of circulating tumor cell separation and enrichment, Insufficient accuracy of detection results and other problems, to achieve the effect of fast automatic separation and purification, high enrichment efficiency and purity, and shorten detection time

Active Publication Date: 2017-11-28
SUREXAM BIO TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These techniques have effectively achieved the separation and enrichment of circulating tumor cells in biological fluid samples to a certain extent, but due to the fact that the physical properties (such as size, density, and electrochemical properties) of circulating tumor cells overlap with normal blood cells. , and no unified standard circulating tumor cell surface markers have been found yet, and there are high differential expressions of circulating tumor cell surface markers in different states, resulting in false negative and false positive results in the separation and enrichment of circulating tumor cells, resulting in Insufficient accuracy of test results

Method used

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  • Circulation tumor cell separation and enrichment micro-fluidic chip and enrichment method thereof
  • Circulation tumor cell separation and enrichment micro-fluidic chip and enrichment method thereof
  • Circulation tumor cell separation and enrichment micro-fluidic chip and enrichment method thereof

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Embodiment 1

[0033] see figure 1 As shown, this embodiment provides a microfluidic chip 10 for separating and enriching circulating tumor cells. A circulating tumor cell separation and enrichment microfluidic chip 10 includes a sample processing device and a separation and purification chamber 200 .

[0034] The sample processing device has a sample processing chamber 100 and a sample processor 120 . The sample processor 120 is provided in the sample processing chamber 100 . The sample processor 120 has an ellipsoidal structure. The sample processor 120 is used to remove the red blood cells in the sample, and make the white blood cells 30 in the sample hybridize with the antibody on the magnetic beads.

[0035] The sample processing chamber 100 has a sample inlet 110 and a sample outlet 140 , two ends of the sample processor 120 are respectively connected to the sample inlet 110 , and one end of the microchannel 210 is connected to the sample outlet 140 . The sample processing device a...

Embodiment 2

[0053] Enrichment and identification of circulating tumor cells in a sample by using the circulating tumor cell separation and enrichment microfluidic chip 10 in Example 1

[0054] 1. Separation and purification

[0055] 1. Sample injection: 20 peripheral blood samples (5 mL each) of tumor patients were injected into the sample processor 120 through the injection port 110 and marked.

[0056] 2. Processing by the sample processor 120: after the sample injection is completed, the plasma in the sample is removed by the sample processor 120 . Add 2 mL of erythrocyte lysate into the sample processor 120, lyse at room temperature for 5 minutes, remove the supernatant, and wash twice with 1 mL of PBS. Finally, 500 μL of magnetic beads embedded with antibodies were added to the sample processor 120, and the hybridization reaction was performed for 30 minutes.

[0057] 3. Separation and enrichment of circulating tumor cells Microfluidic chip 10 Separation and purification: Open the ...

Embodiment 3

[0068] Using the circulating tumor cell separation and enrichment microfluidic chip 10 in Example 1 to detect cell lines

[0069] 1. Selection of cell lines

[0070] In this example, the epithelial cell line MCF-10A, the mesenchymal tumor cell line U118, the epithelial-mesenchymal mixed lung cancer cell line PC-9 and the negative control CCRF-HSB-2 lymphoblastoid cells were used for the experiment. Those skilled in the art can obtain it by purchasing as long as they know the name of the cell line. A certain amount of the above-mentioned cells was collected with a cell counter, and added to 5 mL of normal human peripheral blood to prepare samples with cell concentration gradients as shown in Table 2.

[0071] Table 2 Sample Cell Concentration

[0072]

[0073]

[0074] 2. Sample testing

[0075] Using the microfluidic chip 10 for separating and enriching circulating tumor cells in Example 1 and the method for separation, purification and identification in Example 2, sa...

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Abstract

The invention discloses a circulation tumor cell separation and enrichment micro-fluidic chip and an enrichment method thereof. The circulation tumor cell separation and enrichment micro-fluidic chip comprises a sample treatment chamber and a separation purification chamber, wherein the separation purification chamber comprises a micro channel, a magnetic field adsorption part, a magnetic collection chamber, a cell collection chamber and a filtering membrane; a sub-channel is formed inside the micro channel; the magnetic field adsorption part is arranged on the inner wall of the sub-channel; one end of the micro channel is communicated with the sample treatment chamber; the other end of the micro channel is communicated with the magnetic collection chamber and the cell collection chamber in sequence; the magnetic collection chamber is used for collecting magnetic beads which are not adsorbed by the inner wall of the sub-channel; the filtering membrane which is capable of retaining circulation tumor cells is arranged inside the cell collection chamber; and a liquid outlet from which a liquid filtered by the filtering membrane flows out is formed in the cell collection chamber. The circulation tumor cell separation and enrichment micro-fluidic chip is efficient, rapid, good in specificity and high in accuracy.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a microfluidic chip for separation and enrichment of circulating tumor cells and an enrichment method thereof. Background technique [0002] Circulating tumor cells (CTCs) were first discovered in 1869. They originate from primary tumors or metastatic tumors. They acquire the ability to escape from the basement membrane and enter blood vessels by invading the tissue matrix. Generic term for tumor cells. The current theory of tumor metastasis holds that the formation of CTCs is a sign of distant metastasis of tumors. The content of circulating tumor cells in human blood cells is very small, and only about 1-10 CTCs are contained in 1 mL of blood. Therefore, its detection and counting methods must be efficient, fast, specific, sensitive and accurate. Currently, CTC isolation and enrichment methods are mainly divided into non-specific enrichment methods and specific enrichment methods....

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/574
CPCG01N33/574
Inventor 刘志明吴诗扬廖传荣
Owner SUREXAM BIO TECH
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