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Microfluidic chip based on nanometer drug delivery system screening 3D solid tumor model and preparation method and application thereof

A microfluidic chip and three-dimensional entity technology, which is applied in drug screening, biochemical equipment and methods, compound screening, etc., can solve the problems of accuracy and reliability of analysis results, simple models, and poor integration of drug screening, etc., to achieve easy The effect of mass production, high-throughput detection, and miniaturization

Active Publication Date: 2017-11-21
SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0008] In summary, the introduction of the international cutting-edge microfluidic chip technology into the drug efficacy evaluation research of nano-medicine can overcome the simple model, poor integration of drug screening, and poor accuracy and reliability of analysis results in conventional evaluation and analysis methods. And other issues

Method used

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  • Microfluidic chip based on nanometer drug delivery system screening 3D solid tumor model and preparation method and application thereof
  • Microfluidic chip based on nanometer drug delivery system screening 3D solid tumor model and preparation method and application thereof
  • Microfluidic chip based on nanometer drug delivery system screening 3D solid tumor model and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0074] Embodiment 1 The structure of the chip of the present invention

[0075] The chip of the present invention is composed of a bonded PDMS substrate and a cover sheet. Its structure is as figure 1 As shown, the structure of the cover sheet is three parallel channels, and the parallel channels are connected by connecting channels. Among them, including drug perfusion and single-layer blood vessel formation microchannels (1), connecting channels of extracellular matrix (2), and microchannels with 14 "U" groove structures formed by three-dimensional tumor multicellular spheroids (3) ; The substrate has a shallower parallel channel that facilitates cell capture. The microtissue formed by the present invention is very similar to the microenvironment of the solid tumor in the body, and the penetration of the drug in the microtissue can be observed in real time. Its "U" structure corresponds to each well of a 96-well plate, which is conducive to accurate and rapid drug delivery...

Embodiment 2

[0076] Embodiment 2 takes PDMS as the chip of the present invention of substrate and cover sheet to make

[0077] (1) Put the silicon wafer into concentrated H 2 SO 4 / H 2 o 2 (3 / 1) The solution was boiled slightly for 30 minutes, and after cooling, it was taken out and rinsed with deionized water until it became neutral. After ultrasonication of ethanol for 5 minutes, it was heated on a heating plate for 30 minutes.

[0078] (2) Spin and coat a thin layer of SU-8 photoresist on the silicon wafer, and make the thickness of the glue layer about 50 μm by controlling the speed of the spin. will have figure 2 The mask of the structure is attached to the silicon wafer, and the ultraviolet light passes through the photomask to expose the photoresist. On the silicon wafer, the thickness of the glue is about 50 μm, which will have image 3 The mask of the structure is bonded to the silicon wafer, and the ultraviolet light is used for secondary exposure, and the unexposed part i...

Embodiment 3

[0081] Example 3 Using the chip prepared in Example 2 to form a three-dimensional solid tumor microtissue

[0082] (1) if figure 1 , the channel (2) of the chip obtained by the present invention is perfused with basement membrane extract (BME) gel to simulate the extracellular matrix, and then the channels (1) and (3) are modified with 1% (w / v) polyvinyl alcohol (PVA) ), to prevent cell adhesion to PDMS.

[0083] (2) Set the concentration to 1×10 5 Tumor cells / mL are poured into the channel (3), so that the cells are uniformly and effectively captured in the "U"-shaped groove, and uniform tumor multicellular microspheres are formed after 48 hours of culture.

[0084] (3) Set the concentration to 5×10 4 Each / mL human umbilical vein endothelial cells (HUVECs) flowed into the channel (1), and the chip was placed vertically to allow the cells to grow on the BME glue. After 2 hours, the chip was placed horizontally to remove excess cells in the channel. Because BME has adhesion f...

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Abstract

The invention discloses a microfluidic chip based on a nanometer drug delivery system screening 3D solid tumor model and a preparation method and the application thereof. The microfluidic chip consists of a substrate and a cover plate which are bonded together, wherein the cover plate structurally comprises three parallel microchannels, which are mutually connected and include a microchannel (1) for drug infusion and monolayer angiogenesis, an extracellular matrix connecting channel (2), and a microchannel (4) formed by three-dimensional tumor multicellular spheres and having many U-shaped groove structures (3); the substrate has a parallel channel (5) which facilitates cell capture. The microfluidic chip can achieve real-time monitoring of targeted delivery of the nanometer drug delivery system and online evaluation of drug toxicity at the same time, so that the drug evaluating efficiency is greatly improved; in addition, the microfluidic chip has better tumor simulating capability and provides more accurate data for preclinical drug evaluation; furthermore, the microfluidic chip platform has the advantages of small reagent dosage, low cost, simultaneous analysis of a large number of samples and the like, and has a relatively good drug screening application prospect.

Description

technical field [0001] The invention relates to the technical field of microfluidic chips, in particular to a microfluidic chip for a three-dimensional solid tumor microenvironment for nano drug delivery system screening. Background technique [0002] Malignant tumors are major diseases that seriously threaten human life and health. According to recent data, there are 10 million new cases worldwide each year, and more than 6 million people die of malignant tumors. In 2015, the number of cancer patients in my country was nearly 4.3 million, ranking first in the world, and the mortality rate accounted for 22.3% of the global total, and showed a continuous growth trend [1]. The emergence of nano drug delivery system provides a new gospel for the treatment of malignant tumors. The nano drug delivery system can effectively solve the problems of poor water solubility and easy degradation during the transportation process in the body. It can effectively improve the circulation ti...

Claims

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Application Information

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IPC IPC(8): B01L3/00C12M1/34C12M3/00
CPCB01L3/502707B01L2300/0887B01L2300/12C12M23/12C12N2503/02G01N33/5011G01N2500/10
Inventor 蒋宇扬高丹陈永丽
Owner SHENZHEN GRADUATE SCHOOL TSINGHUA UNIV
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