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A biomimetic coating of red blood cells for enrichment of circulating tumor cells

A technology for tumor cells and red blood cells, applied in the field of red blood cell bionic coatings, can solve problems such as unsatisfactory effects, achieve firm and more efficient capture, and avoid non-specific adsorption effects

Active Publication Date: 2022-01-04
珞珈聚芯(珠海横琴)科技服务中心(有限合伙)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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  • A biomimetic coating of red blood cells for enrichment of circulating tumor cells
  • A biomimetic coating of red blood cells for enrichment of circulating tumor cells
  • A biomimetic coating of red blood cells for enrichment of circulating tumor cells

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Embodiment 1

[0051] The features and advantages of the present invention can be further understood through the following detailed description in conjunction with the accompanying drawings. The examples provided are only illustrative of the method of the present invention and do not limit the rest of the present disclosure in any way. [Example 1] Biomimetic coating of red blood cells prepared by using polylysine to adhere to glass slides

[0052] Modification process of red blood cells: Mix 30 μL of fresh red blood cells from healthy people with 100 μL of DSPE-PEG-FA (1 mg / mL) in PBS, incubate at 4°C for 30 minutes, and centrifuge and wash three times with PBS to obtain folic acid-modified red blood cells. Then mix the erythrocytes with surface-modified folic acid and 10 mg / mL polybrene PBS solution, incubate at 4°C for 30 minutes, wash with PBS for three times to obtain erythrocytes with surface-modified polybrene and folic acid, and disperse them in PBS ready in solution.

[0053] Prep...

Embodiment 2

[0054] [Example 2] Erythrocyte bionic coating captures MCF-7 cells in PBS

[0055] 10 μL of MCF-7 cell suspension (density 5×10 5 cells / mL), dropwise added to the prepared red blood cell biomimetic coating and incubated at 4°C for 120min, and washed with PBS three times to wash away the uncaptured MCF-7 cells. Field photos such as image 3 shown.

[0056] Evaluation of the efficiency of biomimetic coatings on erythrocytes to capture MCF-7 cells in PBS. A total of three groups of experiments were carried out, and each group contained five samples. Five prepared functionalized red blood cell bionic coatings were prepared, and 10 μL, 20 μL, 30 μL, 40 μL, and 50 μL of pre-stained with FDA were dropped on each red blood cell bionic coating. MCF-7 cell suspension (density 5×10 5 cells / mL), which is equivalent to adding 5,000, 10,000, 15,000, 20,000, and 25,000 MCF-7 cells. After the samples of each group were mixed evenly, they were incubated at 37°C for 120 minutes. After the ...

Embodiment 3

[0057] [Example 3] Evaluation of the anti-leukocyte adhesion ability of red blood cell bionic coating

[0058] FDA staining was performed on the extracted buffy coat cells. Add it dropwise on the prepared functionalized erythrocyte biomimetic coating, incubate at 37°C for 2 hours, take 1mL of PBS to wash the biomimetic coating, and take fluorescent photos of the biomimetic coating before and after cleaning with a fluorescence microscope. The results are as follows: Figure 5 shown. It shows that white blood cells cannot adhere to the biomimetic coating of red blood cells at all, and the biomimetic coating of red blood cells has a strong ability to resist adhesion of white blood cells.

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Abstract

The invention discloses a red blood cell bionic coating for enriching circulating tumor cells. Through the dense arrangement of red blood cells on the substrate, non-specific adsorption of white blood cells on the substrate surface is avoided, and at the same time, folic acid (FA) or antibodies modified on the surface of red blood cells can directly target CTCs. The functionalized red blood cell bionic coating relies on its fluidity On the surface of the cell membrane, circulating tumor cells can be captured more firmly and efficiently, with a capture efficiency of more than 90%, and a CTCs capture purity of more than 80%. Red blood cells are derived from the same donor, avoiding the introduction of foreign matter, and high-purity CTCs can be isolated by using red blood cell lysate.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a red blood cell bionic coating for enriching circulating tumor cells. Background technique [0002] Circulating tumor cells (CTCs) are cancer cells that circulate in the bloodstream after shedding from a primary or metastatic tumor. It can travel to distant organs through the blood, triggering cancer metastasis and causing 90% of cancer-related deaths. Isolation and analysis of these CTCs in peripheral blood (liquid biopsy) has attracted attention because of its significance for early cancer diagnosis, treatment monitoring, prognosis assessment, and metastasis diagnosis. However, the number of CTCs in peripheral blood is extremely small, usually only single digits of CTCs in one milliliter of blood, which poses a great challenge to the separation and enrichment of CTCs. [0003] An excellent method for isolating and enriching CTCs should have extremely high sen...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12N5/09
CPCC12M23/02C12M25/02C12N5/0694C12N2509/00
Inventor 刘威张陶冶
Owner 珞珈聚芯(珠海横琴)科技服务中心(有限合伙)
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