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Device and method for culturing high-throughput organoids by using micro-array deep well

A culture device and microarray technology, applied in the field of in vitro culture and analysis of organoids, can solve the problems of relying on operating techniques, low efficiency of organoid formation, and uneven size of organoids cultured in vitro, so as to achieve good application prospects and improve the number of inconsistencies , The effect of improving the efficiency of organoid formation

Active Publication Date: 2021-01-15
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] In order to overcome the above problems, the present disclosure provides a device and method for cultivating high-throughput organoids using microarray deep wells to solve the problem of inhomogeneous in vitro culture of existing organoids, which relies heavily on operating techniques and type organoids. Problems such as low organ formation efficiency

Method used

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  • Device and method for culturing high-throughput organoids by using micro-array deep well
  • Device and method for culturing high-throughput organoids by using micro-array deep well
  • Device and method for culturing high-throughput organoids by using micro-array deep well

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Embodiment 1

[0074] This embodiment provides a device and method for culturing high-throughput organoids using microarray deep wells, wherein the device is used for culturing primary mouse small intestine organoids, which specifically includes the following steps:

[0075] 1. Fabrication of polydimethylsiloxane microarray deep well microfluidic chip. First, using photolithography technology, a diameter of 200 μm, a height of 200 μm, and a distance between the centers of any two adjacent micropillars of 400 μm are produced. The ratio of polydimethylsiloxane A liquid and coagulant B liquid is 10:1, after fully mixing, degassing in vacuum, pouring on the obtained photolithographic micropillar mold, curing at 85°C for two hours The mold is separated from the polydimethylsiloxane.

[0076] 2. Pretreatment and assembly of polydimethylsiloxane microarray deep well microfluidic chip. Before the polydimethylsiloxane microarray deep-well microfluidic chip is inserted into the cells, the chip surfa...

Embodiment 2

[0081] This embodiment provides a high-throughput culture method for tumor organoids, comprising the following steps:

[0082] 1. Fabrication of polydimethylsiloxane microarray deep well microfluidic chip. First, using photolithography technology, a diameter of 200 μm, a height of 200 μm, and a distance between the centers of any two adjacent micropillars of 400 μm are fabricated. The ratio of polydimethylsiloxane A liquid and B liquid is 10:1, after fully mixing, degassing in vacuum, pouring on the obtained photolithography micro-column mold, curing at 85 °C for two hours, the mold and Dimethicone isolated. Among them, liquid A is polydimethylsiloxane PDMS liquid, and liquid B is a coagulant.

[0083] 2. Pretreatment and assembly of polydimethylsiloxane microarray deep well microfluidic chips. Before the polydimethylsiloxane microarray deep-well microfluidic chip is inserted into the cells, the chip surface and the bottom surface are dedusted, and the surface dust is remov...

Embodiment 3

[0088] Such as Figure 8 as shown, Figure 8 It is a schematic diagram of drug testing performed according to the microfluidic control system and polydimethylsiloxane microarray deep well microfluidic chip in Example 3 of the present disclosure. This embodiment provides a method for culturing tumor organoids for drug testing, comprising the following steps:

[0089] 1. Fabrication of polydimethylsiloxane microarray deep well microfluidic chip. Firstly, a polydimethylsiloxane microarray deep-well chip template with a diameter of 200 μm, a height of 200 μm, and a center-to-center distance of 400 μm between any two adjacent microcolumns was fabricated using photolithography technology. The ratio of polydimethylsiloxane A liquid and B liquid is 10:1, after fully mixing, degassing in vacuum, pouring on the obtained photolithography micro-column mold, curing at 85 °C for two hours, the mold and Dimethicone isolated. Among them, liquid A is polydimethylsiloxane PDMS liquid, and l...

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Abstract

The invention relates to a device and method for culturing high-throughput organoids by using a micro-array deep well. The device comprises a cell culture substrate, a polydimethylsiloxane micro-arraydeep well micro-fluidic chip and a micro-fluidic control system, wherein the cell culture substrate is bonded under the polydimethylsiloxane micro-array deep well micro-fluidic chip; the polydimethylsiloxane micro-array deep well micro-fluidic chip is bonded under the micro-fluidic control system, and meanwhile is bonded over the cell culture substrate; and the micro-fluid control system is bonded over the polydimethylsiloxane micro-array deep well micro-fluidic chip. According to the device and the method for culturing the high-throughput organoids by using the micro-array deep well, the device comprising the cell culture substrate, the polydimethylsiloxane micro-array deep well micro-fluidic chip and the micro-fluidic control system is employed, has a simple structure, convenient and controllable to operate, and overcomes the defect that sizes of the high-throughput organoids are very hard to be uniform due to uncontrollability of the operation process.

Description

technical field [0001] The present disclosure relates to the technical field of in vitro culture and analysis of organoids, in particular to a device and method for cultivating high-throughput organoids using microarray deep wells. Background technique [0002] Organoids are three-dimensional tissue structures generated from stem cells or progenitor cells that can mimic corresponding in vivo organs in terms of cell type, tissue structure, and function. Organoids are generally cultured by two-dimensional cell culture methods, which can well control the growth environment of homogeneous cells, facilitate microscopic analysis and observation of medium changes, and maintain cell proliferation of most cell types. The two-dimensional cell culture method provides a large amount of theoretical basis and experimental data for human cell research. [0003] However, most cells in tissues and organs are three-dimensional structures, and they produce physiological functions through the ...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M1/36C12N5/071C12N5/09C12Q1/02B01L3/00
CPCC12M21/08C12M23/16C12M23/20C12M41/00C12N5/0679C12N5/0693C12N5/0631G01N33/5011B01L3/502707C12N2503/02G01N2500/10
Inventor 熊春阳修继冬黄建永
Owner PEKING UNIV
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