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Erythrocyte cluster for enriching circulating tumor cells (CTCs) based on size filtering method

A tumor cell and red blood cell technology, applied in animal cells, vertebrate cells, blood/immune system cells, etc., can solve the problems that the purity of CTCs needs to be improved, and the size difference between white blood cells and CTCs cannot be enlarged.

Active Publication Date: 2020-10-27
SHENZHEN RES INST OF WUHAN UNIVERISTY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these are all bionic designs at the nanometer scale, which cannot enlarge the size difference between white blood cells and CTCs, which leads to the need to improve the purity of CTCs obtained by filtration.

Method used

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  • Erythrocyte cluster for enriching circulating tumor cells (CTCs) based on size filtering method
  • Erythrocyte cluster for enriching circulating tumor cells (CTCs) based on size filtering method
  • Erythrocyte cluster for enriching circulating tumor cells (CTCs) based on size filtering method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] [Example 1] Preparation of normal red blood cell clusters using 5-6μm polystyrene microspheres with surface modified carboxyl groups

[0055] Red blood cell modification process: mix 30μL of healthy human fresh red blood cells with 100μL of 5000-molecular-weight DSPE-PEG-FA (1mg / mL) PBS solution, incubate at 4℃ for 30min, centrifuge and wash with PBS three times to obtain folic acid Modified red blood cells. Then mix the surface-modified folic acid red blood cells with a 10mg / mL polybrene PBS solution, incubate at 4°C for 30 minutes, and wash three times with PBS to obtain surface-modified polybrene and folic acid red blood cells, and disperse them Put it in PBS solution for later use.

[0056] Preparation of red blood cell clusters: Wash the 5-6μm polystyrene microspheres with surface modified carboxyl groups three times with PBS, and then add more than 100 times the number of surface modified polybrene and folic acid red blood cells, and mix them evenly After centrifugat...

Embodiment 2

[0058] [Example 2] Red blood cell clusters or fluorescent red blood cell clusters capture MCF-7 cells

[0059] 1. Red blood cell clusters capture MCF-7 cells in PBS: take 50μL of red blood cell cluster suspension (density 1.2×10 7 Cells / mL), add 10uL of MCF-7 cell suspension (density 5×10 5 Pieces / mL), after mixing, place it at 4°C and incubate it for 120min, blow off the mixture at the bottom of the centrifuge tube after incubation, and filter it through a PDMS porous membrane with a pore size of 20μm to remove excess red blood cell clusters. That is, the MCF-7 cells captured by the red blood cell clusters can be obtained, and the bright field photos taken under the optical microscope are as image 3 (a) Shown.

[0060] 2. Fluorescent red blood cell clusters were captured in PBS to capture MCF-7 cells: 50 μL of the fluorescent red blood cell cluster suspension prepared in Example 1 (density 1.2×10 7 Cells / mL), add 10μL of MCF-7 cell suspension (density 5×10 5 Pieces / mL), after mixi...

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Abstract

The invention discloses an erythrocyte cluster for enriching circulating tumor cells (CTCs) based on a size filtering method, and belongs to the technical field of biological medicine. By designing amicron-level CTCs targeting biomimetic material, erythrocytes are combined with a micro-sphere to obtain an erythrocyte cluster type biomimetic material; the surface of each erythrocyte is modified with polybrene, thereby eliminating original electrostatic repulsion between the erythrocytes, and then, dense full-layer coverage of the erythrocytes on the surface of the micro-sphere can be realized.Wrapping the micro-sphere with the erythrocytes can avoid non-specific adsorption of leukocytes on the surface of the microsphere; meanwhile, folic acid (FA) modifying the surface of erythrocyte cluster or an antibody which can specifically recognize and bind surface biomarkers of the CTCs can directly target the CTCs to achieve increase of sizes of the CTCs, and accordingly, leukocytes in a blood sample of a clinical patient can be finally removed in a manner of filtering, and the high-purity CTCs are obtained efficiently.

Description

Technical field [0001] The invention belongs to the technical field of biomedicine and mainly relates to a red blood cell cluster based on a size filtration method to enrich circulating tumor cells. Background technique [0002] The local or regional recurrence and distant metastasis of tumors pose a severe test for the early diagnosis and cure of tumors. In recent years, molecular biology and clinical studies have shown that circulating tumor cells (CTCs) lead to tumor invasion and micrometastasis. CTCs are formed by the original tumor cells shed from the primary tumor and enter the blood system of the human body. After the circulation and diffusion of the peripheral blood, CTCs eventually metastasize to other tissues and organs of the human body, and then form the same type of tumor as the original site. The secondary tumors caused the metastasis and recurrence of tumors and cancers. By capturing the circulating tumor cells in the peripheral blood and analyzing the changes in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/078G01N33/569
CPCC12N5/0641G01N33/56966
Inventor 刘威彭伟
Owner SHENZHEN RES INST OF WUHAN UNIVERISTY
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