Micro-fluidic chip as well as construction method and application thereof

A microfluidic chip and microchannel technology, applied in the field of biomedicine, can solve the problems of high time cost, low efficiency, unsatisfactory treatment effect, etc., and achieve the effects of simple preparation method, improved efficiency and simulation effect.

Pending Publication Date: 2022-06-14
SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are still certain defects in the related technology
Naked siRNA is easily degraded in the body, which is not ideal for long-term effective treatment, while the adenoviral vector used in CRISPR / Cas9 technology has certain safety risks, and the efficiency of non-viral vectors is low
At present, there are certain deficiencies in the research on the signaling pathway of tumor-nerve-stellate cell relationship and tumor behavior, and the traditional animal experiment method for screening different drug combinations is inefficient, time-consuming, and has ethical issues

Method used

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  • Micro-fluidic chip as well as construction method and application thereof
  • Micro-fluidic chip as well as construction method and application thereof
  • Micro-fluidic chip as well as construction method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1 2

[0069] Example 1 Construction of a two-dimensional pancreatic cancer-neural-stromal microenvironment model

[0070] 1. Extraction and culture of primary dorsal root neurons

[0071] Isolation and culture of primary dorsal root ganglion (Dorsal root ganglion, hereinafter referred to as DRG): SD rats within 24 hours of birth were decapitated on ice, and their vertebrae were removed, and the spinal cord was exposed using sterile rongeurs. The dorsal root ganglia distributed on both sides of the spinal cord were removed and placed in pre-cooled DMEM (Dulbecco's modified eagle medium) medium (Gibco, product number: 11965092). Rinse the DRG tissue gently with DMEM medium and transfer it to a 15mL centrifuge tube, digest it with 10× digestion solution (Gibco, catalog number: 15090046) at 37°C for 30min, and shake the centrifuge tube every 5min to make the DRG tissue and the digestion solution fully touch. When the digestion is over, stop the digestion with DMEM medium and gently ri...

Embodiment 2 3

[0081] Example 2 Construction of a three-dimensional pancreatic cancer-neural-stromal cell microenvironment model

[0082] Using 3D printing technology, nerve cells, pancreatic cancer cells, and pancreatic stellate cells are wrapped in hydrogel and arranged in parallel. micrometer), cancer cell area (400 microns wide), hydrogel area (400 microns wide), nerve cell area (500 microns wide). The microporous structure in the hydrogel region ensures the mutual transfer of cytokines, drugs, etc.

[0083] The 3D model microenvironment is based on the 2D model microenvironment combined with hydrogel to form a physicochemical microenvironment similar to that in the body, providing microcomponents similar to those in the body for cell growth, and can further restore the microenvironment state of pancreatic cancer in vivo. .

Embodiment 3 3

[0084] Example 3 Application of three-dimensional model microenvironment in nano-drug screening

[0085] 1. Construction of a microenvironment-related nanodrug library based on a 3D model

[0086] Based on the design and synthesis of biomedical nanomaterials accumulated in our laboratory, combined with the synthesis methods of related technologies, we will design and synthesize different types of nanomaterials, including inorganic nanoparticles (such as gold nanoclusters), cationic liposomes (such as phospholipids) ), solid lipid nanoparticles (such as lipid-PLGA), etc.

[0087] In the embodiment of the present invention, solid lipid nanoparticles lipid-PLGA with different particle sizes, different surface charges, different hardness and different surface modifications are controlled and synthesized by microfluidic technology, which is used to construct a supplementary nerve-related nanomedicine library. At the same time, NGF-siRNA, BDNF-siRNA or related CRISPR / Cas9 systems t...

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Abstract

The invention discloses a micro-fluidic chip as well as a construction method and application thereof. The micro-fluidic chip comprises a polydimethylsiloxane layer and a substrate layer, wherein the polydimethylsiloxane layer comprises three or more than three micro-channels; the micro-channels are connected through a plurality of micro-channels which are arranged in parallel; the micro-channel is provided with a pore channel connected with the outside; the substrate layer is at least one of a glass substrate, a polydimethylsiloxane substrate or a polystyrene substrate, the preparation method of the micro-fluidic chip is simple, and the culture of tumor nerve cells can be realized. A pancreatic cancer-nerve-stromal cell microenvironment model constructed by utilizing the micro-fluidic chip disclosed by the invention can realize joint construction of a pancreatic cancer stromal microenvironment and a neural microenvironment, and can also perform qualitative and quantitative evaluation on nano-drugs targeting the neural microenvironment and the stromal microenvironment; the method plays an important role in screening related drugs for treating pancreatic cancer.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a microfluidic chip and a construction method and application thereof. Background technique [0002] In recent years, more and more attention has been paid to the regulation of cancer microenvironment (mainly stromal microenvironment and neural microenvironment), mainly including stellate cells, nerve cells, immune cells, vascular cells and other cells and tumor cells and their secreted insoluble Extracellular matrix and soluble cytokines. The neural microenvironment of pancreatic cancer affects its pathological process, mainly because cancer cells and nerve cells can promote the growth of each other through neurotrophic factor, nerve growth factor (Nerve growth factor, hereinafter referred to as NGF), cytokines and neurotransmitters. cause pancreatic cancer. Second, the connective tissue (stromal) microenvironment can regulate the occurrence and development of pancreatic cancer. Wh...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00B01L3/00C12N5/071C12N5/0793C12N5/09C12Q1/02
CPCC12M23/16C12N5/0676C12N5/0693C12N5/0619G01N33/5011G01N33/5044G01N33/5058B01L3/502707C12N2502/00G01N2500/10C12N2513/00C12N2503/02
Inventor 蒋兴宇刘晓艳田瑞军吴德成
Owner SOUTH UNIVERSITY OF SCIENCE AND TECHNOLOGY OF CHINA
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