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Integrated microfluidic chip and its method for three-dimensional tumor localization, construction and recovery

A microfluidic chip, three-dimensional technology, applied in the direction of tumor/cancer cells, biochemical equipment and methods, enzymology/microbiology devices, etc., can solve the problem of three-dimensional tumor construction and analysis, recovery operation and Post-analysis research, unfavorable 3D tumor operation, 3D tumor structure destruction, etc., to achieve good time and space control and flexibility, simple and fast operation, and precise cell positioning

Active Publication Date: 2016-07-06
NORTHWEST A & F UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the developed chip microfunctional units, such as microsieves and micropores, can capture tumor cells in situ, they cannot realize real-time space-controlled tumor cell localization, three-dimensional tumor construction and analysis, and recovery operations due to their fixed structures. and post-analytic studies
In addition, if extreme conditions are used for sample recovery, such as the use of extremely high flow rates, it will inevitably cause the destruction of the three-dimensional tumor structure and the residue of tumor cells in these fixed micro-functional units, which is not conducive to three-dimensional tumor manipulation.

Method used

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  • Integrated microfluidic chip and its method for three-dimensional tumor localization, construction and recovery
  • Integrated microfluidic chip and its method for three-dimensional tumor localization, construction and recovery
  • Integrated microfluidic chip and its method for three-dimensional tumor localization, construction and recovery

Examples

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Effect test

preparation example Construction

[0034] The preparation material of the integrated microfluidic chip of the present invention is polydimethylsiloxane (PDMS), which is irreversibly sealed with glass or transparent plastic material coated with PDMS polymer, thereby ensuring the quality of the microfluidic chip used and The integrity of each micro-functional unit in the chip.

[0035] The heights mentioned in the present invention all refer to the height in the working chamber.

[0036] The integrated microfluidic chip provided by the invention is used for high-throughput tumor cell localization, three-dimensional tumor bionic construction and tumor recovery, and the cell samples are mammalian tumor cells and tumor cell lines.

[0037] The invention provides an operation method for high-throughput tumor cell localization, three-dimensional tumor bionic construction and tumor recovery based on an integrated microfluidic chip. The anti-cell adhesion solution is injected into the microcavity from the inlet of the ...

Embodiment 1

[0043] The integrated microfluidic chip used in this example was designed and manufactured by the applicant's laboratory. The material used to prepare the integrated microfluidic chip is PDMS polymer, which is irreversibly sealed on the glass surface coated with PDMS film.

[0044] The integrated microfluidic chip used in this example consists of a sample flow layer and a pneumatic control layer.

[0045] Such as figure 1 As shown, the sample flow layer contains 1 inlet (1), 8 microcavities, 8 outlets (2 and 3) and 1 symmetrical micropipe network. Each microcavity is correspondingly connected to one outlet (2 and 3), and the eight microcavities are symmetrically distributed and connected to the inlet (1) by a symmetrical micropipe network.

[0046] The pneumatic control layer contains 1 air inlet (4), 360 pneumatic control structures and 1 closed-end micro-pipeline network. The height of the sample flow layer is 100 μm, the height of the pneumatic control structure is 30 μm...

Embodiment 2

[0053] This example is the high-throughput HepG2 liver cancer cell location capture and three-dimensional tumor construction of the microfluidic chip provided in Example 1:

[0054] Firstly, anti-cell adhesion modification was performed on the microfluidic chip described in Example 1, that is, bovine serum albumin (BSA) solution with a concentration of 2 mg / mL was injected from the fluid layer inlet of the chip at a flow rate of 5 μL / min through a microsyringe pump (1 ), BSA enters the microcavity through symmetrical microchannels; incubate at room temperature for 2 hours; then perfuse and wash with fresh DMEM cell culture medium;

[0055] Perform cell perfusion on the microfluidic chip described in Example 1, that is, the cell density is 1×10 5 Each / mL liver cancer cell suspension was introduced from the fluid layer inlet (1) of the chip at a flow rate of 5 μL / min, and the cells entered the cell microcavity through symmetrical microchannels ( Figure 5 shown);

[0056] Give...

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Abstract

The invention discloses an integrated microfluidic chip and a three-dimensional tumor location, construction and recycling method. The disclosed chip is composed of a sample flow layer and a pneumatic control layer; the sample flow layer is provided with an inlet, micro-cavities, outlets and a micro pipeline network, and the pneumatic control layer is provided with a gas inlet, pneumatic control structures and an end-closed micro pipeline network. The pneumatic control structures are connected with the gas inlet through the end-closed micro pipeline network. The height of the pneumatic control structures is greater than that of the end-closed micro pipeline network. Tumor cell accurate locating capture, three-dimensional tumor forming and online analysis and recycling operations can be continuously completed in the single microfluidic chip. The operations have extremely strong time and space control and flexibility, and are convenient for multidisciplinary research scholars to master; and the chip has the characteristics of simple and fast operations, low sample consumption, accurate cell location, high efficiency, high throughput and the like, and is suitable for construction and analysis operations on various three-dimensional tumors.

Description

technical field [0001] The invention relates to the technical field of microfluidic chips, in particular to an integrated microfluidic chip and its use in real-time controlled high-throughput tumor cell localization, three-dimensional tumor tissue construction and recovery. Background technique [0002] Cancer is a disease caused by the uncontrolled division and abnormal growth of cells, often accompanied by the formation of malignant tumors, which in turn leads to human and animal disease and eventually death. Extensive tumor research is the driving force for exploring the mechanism of cancer and promoting the progress of clinical treatment. At present, conventional animal in vivo experimental research methods can highly restore the occurrence and development of specific tumors, but lack sufficient advantages in dynamic detection, microenvironment control, and real-time operation and analysis of tumor tissues. In comparison, conventional in vitro tumor cell culture methods...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M1/00C12M1/04C12N5/09
CPCC12M23/16C12M47/04C12N5/0693
Inventor 刘文明王进义王建春许娟李天保涂琴王垚磊
Owner NORTHWEST A & F UNIV
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