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Microfluidic cell array chip for high-throughput medicament screening, method and use

A cell array and microfluidic technology, applied in the direction of supporting/immobilizing microorganisms, biochemical equipment and methods, specific-purpose bioreactors/fermenters, etc., can solve the problem of difficult liquid distribution and operation of micro samples, reagents Large volume, high precision requirements, etc., to achieve the effect of solving disordered cell distribution and low reagent consumption

Inactive Publication Date: 2011-09-21
SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, there are some limitations in cell-level drug screening based on this method. For example, high-throughput screening is generally carried out through multi-well plates (96-well and 384-well plates), and the amount of reagents consumed is relatively large. Although micro-well plates can reduce the amount of reagents, However, the micro-volume solution is easy to evaporate, and the distribution and operation of micro-sample liquid is difficult and requires high precision. In addition, the equipment for high-throughput screening is bulky and expensive
These largely limit the popularization and application of high-throughput screening

Method used

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  • Microfluidic cell array chip for high-throughput medicament screening, method and use
  • Microfluidic cell array chip for high-throughput medicament screening, method and use
  • Microfluidic cell array chip for high-throughput medicament screening, method and use

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Embodiment 1

[0032] Embodiment 1, the preparation of chip

[0033] The microfluidic chip is manufactured by multiple times of soft photolithography and molding. The chip adopts polydimethylsiloxane (PDMS) material with good light transmission, gas permeability and biocompatibility, which is bonded with conventional glass slides to form a cell culture pool array and its microchannel network structure. The key process of chip making is to make a double-layer PDMS (polydimethylsiloxane) chip by making a fluid channel layer mold of double-layer SU-8 negative photoresist to form a dam structure and thermal bonding. Specific steps are as follows:

[0034] 1) On the cleaned silicon substrate, spin-coat diluted SU8-2005 (MicroChemCorp, MA, USA, diluted with SU8-2025 and cyclopentanone at a volume ratio of 100:53), with a thickness of about 5 μm, Put it on a hot plate and bake at 65°C for 1 minute, then raise the temperature to 95°C and keep it for 2 minutes, then slowly cool down to room tempera...

Embodiment 2

[0039] Embodiment 2, the application of chip

[0040] The cells cultured on the chip were human liver cancer cells SMMC-7721, normal human liver cells HL-7702 and umbilical vein endothelial cells. The cell culture medium is RMPI1640 (Gibco), adding 10% fetal bovine serum (Hangzhou Sijiqing) and 1% streptocyanine double antibody (Hangzhou Jinuo Biomedical Technology Co., Ltd.), and the cell digestion solution is 0.25% trypsin and 0.02% EDTA (Hangzhou Jinuo Biomedical Technology Co., Ltd.).

[0041] The chip was irradiated with 75% ethanol and irradiated with ultraviolet light for 1 hour for disinfection. First, use a syringe pump to pass through the cell culture solution into the chip for 2 hours to eliminate the residual ethanol and air bubbles in the chip, which is conducive to the adherent growth of cells in the culture chamber. The three types of cells are respectively introduced into each columnar cell culture chamber of the chip through a syringe pump for cell culture. ...

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Abstract

The invention relates to a microfluidic cell array chip for high-throughput medicament screening, a preparation method thereof and use thereof. The microfluidic cell array chip for high-throughput medicament screening is characterized by sequentially comprising a valve-controlled channel layer, a fluid channel layer and a glass layer suitable for the anchorage growth of cells from top down. The chip can be used in high-throughput medicament screening by the parallel action of different medicament concentrations on various cells. The chip is manufactured by a multixposure one-development Su-8 negative photoresist process and multilayer PDMS bonding and has a structure with a high depth-to-width ratio. The microfluidic cell array chip provided by the invention can realize low reagent consumption and high-throughput co-culture of various cells, parallel analysis of the stimulation effect of different medicament concentrations on different cells and the real-time observation and detectionof a slide of the cells and provide a fire-new technical platform and a method for high-throughput medicament screening and cell-medicament researches.

Description

technical field [0001] The invention relates to a microfluidic cell array chip for high-throughput drug screening, a preparation method and an application, and belongs to the technical field of microfluidic chips. Background technique [0002] High-throughput drug analysis technology is mainly used in the discovery stage of drug research, which is also the basis and key of drug research. Since the 1990s, due to the development of modern technology, high-throughput screening based on 96-well plates is suitable for preliminary screening due to its fast and efficient characteristics. It has been widely used by most international drug research institutions and has been rapidly It has become one of the main technical means for drug screening. However, there are some limitations in cell-level drug screening based on this method. For example, high-throughput screening is generally carried out through multi-well plates (96-well and 384-well plates), and the amount of reagents consu...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/00C12M1/36C12Q1/02
CPCC12M25/06C12M23/04C12M41/36C12M23/20
Inventor 金庆辉郑允焕吴建璋邵建波赵建龙
Owner SHANGHAI INST OF MICROSYSTEM & INFORMATION TECH CHINESE ACAD OF SCI
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