Cancer cell membrane embedded human bone micro-fluidic chip and preparation method and application thereof in separating circulating tumor cells

A microfluidic chip and cancer cell membrane technology, applied in the field of biomedicine, can solve the problems of unfavorable CTC downstream analysis, low cell capture efficiency, low CTC purity, etc., achieve firm and more efficient capture, improve enrichment purity, and low cost Effect

Inactive Publication Date: 2021-11-30
WUHAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the above method releases target cells and non-specifically adhered background cells together, which will lead to low purity of enriched CTCs, which is not conducive to downstream analysis of CTCs.
There are also techniques for recovering single cells from substrates, such as laser cutting, micromanipulation, flow cytometry, etc. However, the existing single cell isolation methods often require the use of external equipment, which is costly and has low cell capture efficiency

Method used

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  • Cancer cell membrane embedded human bone micro-fluidic chip and preparation method and application thereof in separating circulating tumor cells
  • Cancer cell membrane embedded human bone micro-fluidic chip and preparation method and application thereof in separating circulating tumor cells
  • Cancer cell membrane embedded human bone micro-fluidic chip and preparation method and application thereof in separating circulating tumor cells

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preparation example Construction

[0041] According to a typical embodiment of the present invention, a method for preparing a human bone microfluidic chip embedded in a cancer cell membrane is provided, such as Figure 8 As shown, the method includes:

[0042] Step S101, culturing the cell suspension of the cancer cell line, washing and fixing the cancer cells after they attach to the wall, to obtain the cancer cell membrane substrate;

[0043] In this embodiment, the cancer cell lines should be selected from those cancer cell lines that are concentrated in research and have a high degree of postoperative metastasis and recurrence. The following cancer cell lines can be used but are not limited to: breast cancer cell lines, prostate cancer cell lines, colon cancer cell lines, etc. Cancer cell lines, liver cancer cell lines, gastric cancer cell lines, lung cancer cell lines. The breast cancer cell line (ie, MCF-7 cells) is preferably recommended as a tumor cell model for extracorporeal circulation. Because br...

Embodiment 1

[0073] Example 1 Human bone microfluidic chip embedded in cancer cell membrane and its preparation method

[0074] 1. The human bone microfluidic chip MHB-chip embedded in the cancer cell membrane is composed of two functional components, including a polydimethylsiloxane (PDMS) layer with a human bone pattern on the surface and a transparent MCF-7 cell membrane substrate ( MS).

[0075] 2. Preparation method of human bone microfluidic chip embedded in cancer cell membrane

[0076] (1) In order to prepare MS, we will 1×10 6 The MCF-7 cell suspension was added into a culture dish (clean glass was placed in the culture dish) for cultivation. When the cells were fully grown on the glass, the medium was removed and washed with PBS. Next, the cells on the MS were fixed with 2.5% glutaraldehyde, thereby maintaining the nanostructure of the MCF-7 cell membrane. ;

[0077](2) A 10 mg / mL bovine serum albumin solution was introduced into the MCF-7 cell membrane. MCF-7 cell membrane...

Embodiment 2

[0079] The cell membrane photo of the MHB-chip of the embodiment of the present invention is as follows figure 2 shown by figure 2 It can be seen that figure 2 (a) shows that the basal surface is overgrown with MCF-7 cells, figure 2 (b) and figure 2 (c) shows that the prepared substrate surface is a kind of nanostructure. Example 2 Human bone microfluidic chip embedded in cancer cell membrane and its preparation method

[0080] 1. The human bone microfluidic chip MHB-chip embedded in the cancer cell membrane is composed of two functional components, including a polydimethylsiloxane (PDMS) layer with a human bone pattern on the surface and a transparent MCF-7 cell membrane substrate ( MS).

[0081] 2. Preparation method of human bone microfluidic chip embedded in cancer cell membrane

[0082] (1) In order to prepare MS, we will 1×10 6 The MCF-7 cell suspension was added into a culture dish (clean glass was placed in the culture dish) for cultivation. When the cells...

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Abstract

The invention discloses a cancer cell membrane embedded human bone micro-fluidic chip and a preparation method and application thereof in separating circulating tumor cells. The method comprises the following steps of: culturing a cell suspension of a cancer cell strain, washing and fixing cancer cells after the cancer cells are adhered to the wall to obtain a cancer cell membrane substrate; adding a BSA solution and an NHS-S-S-Biotin solution for incubation, soaking the cancer cell membrane substrate in a streptavidin solution, washing with PBS, and adding a biotinylated anti-EpCAM antibody solution for incubation to obtain an EpCAM antibody modified cancer cell membrane substrate; and integrating the EpCAM antibody modified cancer cell membrane substrate with human bone PDMS to obtain a cancer cell membrane embedded human bone micro-fluidic chip , which is called MHB-chip for short. The chip can be used for specifically capturing and releasing trace CTCs in a simulated blood sample and a blood sample of a clinical patient in vitro, and is high in capturing efficiency and high in capturing purity.

Description

technical field [0001] The invention relates to the technical field of biomedicine, in particular to a human bone microfluidic chip embedded in a cancer cell membrane, a preparation method thereof, and an application in separating circulating tumor cells. Background technique [0002] Circulating tumor cells are tumor cells shed from solid tumors into the peripheral blood. Circulating tumor cells will spread to other locations along with the peripheral blood. The metastasis and proliferation of cancer cells is the main reason for the death of tumor patients. Circulating tumor cells carry all the genetic information of tumor patients and can be used as one of the important means of liquid biopsy, which will provide an important basis for patient diagnosis, personalized treatment and postoperative evaluation. However, knowing that there may be only a few CTCs per milliliter of blood, there is an urgent need to develop techniques to isolate rare circulating tumor cells from blo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01L3/00C12N5/09
Inventor 刘威陈贝
Owner WUHAN UNIV
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