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Cell replicated surface and application thereof

A technology of cells and rare cells, which is applied in the field of biofunctional materials, analysis and detection, can solve the problems of the preparation method and application of cell replica surface that have not been reported, and achieve the effect of low preparation cost, high capture rate and high capture rate

Active Publication Date: 2018-09-18
DALIAN UNIV OF TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation method and application of this kind of cell replica surface have not been reported yet.

Method used

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  • Cell replicated surface and application thereof
  • Cell replicated surface and application thereof
  • Cell replicated surface and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] The template cells used in this example are human breast cancer MCF-7 cells (Guangzhou Saiku Biotechnology Co., Ltd.), and the soft engraving material is polymer polydimethylsilane (PDMS); by examining the cell engraved PDMS surface and planar surface Taking the capture of cancer cells as an example, the capture system of the present invention is further described and verified, including the following steps:

[0027] 1) Preparation of cell replica PDMS surface:

[0028] The template cell MCF-7 cells were used at 25000cells / cm 2 The density was seeded in a Petri dish and placed in 37 °C CO 2 In the cell culture incubator, after 24 hours, fix with 4% mass concentration paraformaldehyde solution for 20 minutes, and the fixed cells are dehydrated in absolute ethanol for 30 minutes and dried to obtain the template cell surface. Then, the PDMS prepolymer was poured on the surface of the template cells, and after thermosetting and cross-linking at 80°C for 6 hours, the cells...

Embodiment 2

[0038] In this example, the surface of cell reprinted PDMS is selected, and the substrate material is connected with anti-EpCAM specific antibody by using biotin / bioavidin interaction; by investigating the capture efficiency of cell reprinted PDMS surface on different types of cancer cells, the present invention The capture system for further elaboration and verification, including the following steps:

[0039] 1) The preparation of the cell replica PDMS surface is the same as in Example 1:

[0040] 2) Bioaffinity linking anti-EpCAM specific antibody:

[0041] Treat the surface of PDMS with ultraviolet and ozone for 10 minutes to make it hydrophilized, then place it in a cell culture plate, add n-octadecyltriethoxysilane solution (C18, diluted with ethanol) with a mass concentration of 4%, After reacting at room temperature for 45 minutes, they were washed once with ethanol and water respectively, and washed three times with phosphate buffer saline (PBS). Add 0.5 mg / mL bioti...

Embodiment 3

[0050] In this example, the anti-EpCAM specific antibody cells modified by chemical coupling method were selected as the capture system, and different numbers of MCF-7 cells were added to the whole blood as the artificial blood sample model of CTCs to be captured; and the captured Cancer cells and leukocytes are observed by scanning electron microscope, and the capture system of the present invention is further elaborated and verified, including the following steps:

[0051] 1) The preparation method of the cell replicated PDMS surface is the same as that in Example 1.

[0052] 2) Chemical coupling method to connect anti-EpCAM specific antibody:

[0053] The surface of the cells replicated PDMS was treated with ultraviolet ozone for 5 minutes, put into the cell culture plate, added n-octadecyltriethoxysilane ethanol solution with a mass concentration of 4%, soaked at room temperature for 45 minutes, washed once with absolute ethanol, Rinse once with water and blow dry with ni...

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Abstract

The invention provides a cell replicated surface and an application thereof. The cell replicated surface is a micro-nano topological structure surface used for highly purely capturing rare cells in blood and complementary to a template cell structure, the surface also can be modified with a rare cell specific recognition antibody, and the surface and the antibody have synergistic effects to realize high-efficiency specific capturing. The cell replicated surface has the advantages of simple production technology, low production cost, very high capturing efficiency on the rare cells, superior specificity, and broad-spectrum capturing property to different rare cell types, is especially suitable for capturing rare cells enriched in patient bodies, especially circulating tumor cells, and has agreat guiding effect on cancer prognosis, early diagnosis, curative effect assessment, recurrence prediction and individual treatment.

Description

technical field [0001] The invention belongs to the technical field of biofunctional materials and analysis and detection, and in particular relates to the technology of forming a cell replica surface on the material surface and capturing rare cells in blood, especially circulating tumor cells. Background technique [0002] Some cells in the blood are present in very small amounts but are important for understanding disease, such as stem cells, circulating endothelial cells, residual disease cells, and circulating tumor cells. Accurate detection and analysis of these rare cells is critical to understanding disease processes and pathogenesis, especially circulating tumor cells are closely related to early diagnosis of cancer. Studies have shown that 90% of cancer deaths are caused by cancer metastasis. Circulating tumor cells (CTCs) refer to tumor cells that shed from solid tumor lesions and enter the peripheral blood after undergoing epithelial-mesenchymal transition (EMT). ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09
CPCC12N5/0693C12N2509/00
Inventor 韩璐璐贾凌云孙贺杨立为
Owner DALIAN UNIV OF TECH
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