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A CTC capture and purification substrate based on chitosan nanoparticles and its preparation method

A technology of chitosan nanoparticles and particles, applied in the field of medical clinical CTC separation, to achieve the effect of increasing the number, strong practicability, and good biological activity

Active Publication Date: 2019-02-01
SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, natural biopolymers with better cytocompatibility, including chitosan, have not been applied to the research of CTC capture based on nanoparticle interface.

Method used

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  • A CTC capture and purification substrate based on chitosan nanoparticles and its preparation method
  • A CTC capture and purification substrate based on chitosan nanoparticles and its preparation method
  • A CTC capture and purification substrate based on chitosan nanoparticles and its preparation method

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Embodiment 1 The specific steps of the preparation method of the CTC capture and purification substrate based on chitosan nanoparticles are as follows:

[0064] a) Using low molecular weight chitosan in an organic acid system, after stirring for 4-8 hours at room temperature, a homogeneous transparent chitosan solution is obtained.

[0065] b) electrospray the chitosan solution in step a for 0.1-4h in an electrospinning device, and prepare chitosan nanoparticles on the surface of the glass substrate. Chitosan nanoparticles surface P1, P2, P3, P4, P5 with different nanometer particle size and density were prepared by adjusting the properties of electrospray liquid, electrospray parameters and electrospray time. The surface structure and morphology of prepared different chitosan nanoparticles are shown in figure 2 .

[0066] c) Place the surface of chitosan nanoparticles in step b in ammonia water / methanol solution (the volume ratio of ammonia water to methanol is 4:96...

Embodiment 2

[0070] Example 2 Using the EpCAM-positive breast cancer cell line MCF-7 as a model cell, the capture behavior of P4 on the nanostructured surface to cancer cells was investigated. Figure 4 Different capture behaviors of MCF-7 cells by P4 surface with three different modified interfaces. The surface modified by PEG-aptamer ( Figure 4 , a)) the maximum amount of cell capture, while the PEG-modified surface ( Figure 4 , c)) The amount of cell adhesion is minimal.

Embodiment 3

[0071] Example 3 Using the EpCAM-positive breast cancer cell line MCF-7 as a model cell, the capture efficiency and specificity of different micro-nano structured surfaces for cancer cells were systematically investigated. At the same time, a bare interface without any modification and an interface only modified with PEG were constructed on each nanostructured surface as controls. MCF-7 cells that have been cultured for two days and in good growth state were digested and peeled off with 0.25% trypsin, then discarded the trypsin solution, added fresh culture medium to blow the cells evenly, counted the cells, and adjusted the cell suspension to 10 5 / ml. Place the modified nano-substrate in a 24-well plate, inject 1ml of the prepared cell suspension into each well, incubate in the cell culture incubator for 10-60min, wash with PBS for 3-5 times, and use a fluorescence microscope Captured cells were observed and counted. The experimental results showed that the amount of cell ...

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Abstract

The invention discloses a CTC (circulating tumor cell) capture and purification substrate based on chitosan nanoparticles, its preparation method and application. The present invention selects chitosan with good cytocompatibility to build a three-dimensional nanostructure "soft" substrate, introduces anti-adhesion molecules, such as polyethylene glycol (PEG) molecules to reduce non-specific adhesion of cells on the interface, and utilizes CTC affinity And capture molecules, such as epithelial cell adhesion molecule (EpCAM) aptamers to achieve high specific capture of CTC cells, and then based on the differences in the proliferation properties of CTC and blood cells, the purity of target cells is further improved by in situ culture of captured cells. The CTC capture and purification substrate of the present invention has good cytocompatibility, can maintain the activity of captured cells, and the preparation process is simple and convenient, with low cost, and can realize in situ culture and purification of captured cells, and is also easy to implement on a large scale .

Description

technical field [0001] The invention relates to a CTC separation and purification method, in particular to a CTC capture and purification substrate based on chitosan nanoparticles and its preparation method and application, belonging to the technical field of medical clinical CTC separation. Background technique [0002] Circulating tumor cells (CTCs) refer to tumor cells that have shed from the primary tumor and entered the peripheral blood of the human body. Circulating tumor cells enter the blood circulation and migrate to other tissue parts of the body to cause tumor metastasis, also known as disseminated tumor cells. CTC is closely related to cancer metastasis, treatment effect, cancer recurrence, medication guidance and prognosis, and thus is used as an important biomarker for early diagnosis and treatment of cancer metastasis. Research on CTC is expected to elucidate the internal mechanism of cancer metastasis, drug sensitivity and drug resistance, so as to realize i...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N11/10
Inventor 裴仁军孙娜王金娥
Owner SUZHOU INST OF NANO TECH & NANO BIONICS CHINESE ACEDEMY OF SCI
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