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Device for patterning cocultivation of multiple cells, preparation method and use thereof

A co-culture and cell technology, applied in biochemical equipment and methods, tissue cell/virus culture devices, tissue culture, etc., can solve the limited PDMS microfluidic channel preparation technology, exist on the same substrate, and cannot achieve different species Issues such as cell arrangement and density

Active Publication Date: 2013-10-16
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method can only form continuous cell strips, and its arrangement is controlled by microfluidic channels, that is, the area occupied by cells on the gold surface is determined by the size and interval of the channels, which is limited by the preparation of PDMS microfluidic channels. technology
[0006] It can be seen that the disadvantage of cell co-culture at present is that all cells are in the same way, all are single cells or all are multicellular, or are arranged continuously or discontinuously on the substrate, and it is impossible to achieve different types of cells. Specific combinations of permutations and densities exist on the same substrate

Method used

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  • Device for patterning cocultivation of multiple cells, preparation method and use thereof
  • Device for patterning cocultivation of multiple cells, preparation method and use thereof
  • Device for patterning cocultivation of multiple cells, preparation method and use thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0055] Example 1 Multicellular-multicellular co-culture model

[0056] This embodiment is a device in which different types of cells exist in the same substrate in a specific combination of arrangement and density, and is used for culturing cells to establish a multi-cell-multi-cell co-culture model. The device prepared in this example is as figure 1 , wherein a shows a schematic diagram of the device structure before thiol reassembly in the microfluidic channel, and b shows a schematic diagram of the device structure after thiol reassembly in the microfluidic channel.

[0057] 1) The preparation of the polydimethylsiloxane stamp template, the main process is photolithography, that is, using the characteristics of photoresist that can change its properties under ultraviolet radiation to make a photoresist that is completely consistent with the pattern on the designed mask Silicon wafer template, the specific preparation method can be found in Y.Xia, G.Whitesides, Annual Re...

Embodiment 2

[0079] Example 2 Single-cell-multicellular co-culture model

[0080] This example is to prepare another device in which different kinds of cells exist on the same substrate in a specific combination of arrangement and density, and is used for culturing cells to establish a single-cell-multi-cell co-culture model. MDCK adheres to the wall in the channels on both sides, and NIH 3T3 cells adhere to the pattern in the middle channel. Adjust the appropriate cell density so that only one cell can be accommodated on a single pattern.

[0081] The device prepared in this example is as image 3 , wherein a shows a schematic diagram of the device structure before thiol reassembly in the microfluidic channel, and b shows a schematic diagram of the device structure after thiol reassembly in the microfluidic channel.

[0082] Except step 4), 5), 6), 10), 11), all the other are identical with embodiment 1.

[0083] 4) Using photolithography technology, engrave concave microstructure un...

Embodiment 3

[0089] Example 3 .Selective injury model for combinatorially arranged cells on the same substrate

[0090] This example is to prepare a device in which the same kind of cells exist in the same substrate in a specific combination of arrangement and density, and is used to selectively damage cultured cells, to obtain the arrangement of normal cells and damaged cells on the same plane, and to establish a selective damage model .

[0091] Except steps 10) to 11), all the other are identical with embodiment 1, promptly used device is identical with embodiment 1, as figure 1 shown.

[0092] 10) Prepare a suspension solution of MDCK cells (see Xingyu Jiang, PNAS, 2005, 102, 975-978 for specific methods), MDCK (purchased from Union Medical College Hospital), and the cell density is 10 7pcs / ml, then pass MDCK into the left, middle and right three microfluidic channels, and then put it into the cell incubator, at 37°C, with a carbon dioxide volume concentration of 5%, culture for 2 ...

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Abstract

The invention provides a device for patterning cocultivation of multiple cells, a preparation method and a use thereof. The device provided by the invention comprises 1) substrates, wherein a titanium layer and a gold layer are orderly evaporated on the surface and a mercaptan self-assembled area ended by methyl in arrangement of a specific pattern array is attached to the gold layer; and 2) a polydimethylsiloxane seal comprising at least a micro-groove unit, wherein the polydimethylsiloxane seal is adhered to the substrate to form a plurality of micro-flow channels and the substrate in each micro-flow channel can be selectively modified as a) a substrate accelerating cell adhesion or b) a substrate locally accelerating cell adhesion and locally preventing cell adhesion. The device provided by the invention is prepared by re-assembling mercaptan in the channel and different cell combination arrays can be adhered to a same substrate, thereby breaking through the mode that all cell arrangement positions and densities are same in original cell cocultivation method.

Description

technical field [0001] The present invention relates to a cell co-cultivation device applying microfluidic and cell patterning technology to the field of biomedicine, in particular to a device for co-culturing various cells on the same substrate in combination, arrangement and patterning, its preparation method and application. Background technique [0002] Tissues and organs are a complex multicellular environment, and cell-cell and cell-extracellular matrix interactions affect the behavior of cells. The traditional method of simulating a multicellular environment in vitro can be achieved by mixing different types of cells, or planting another type of cell on a layer of cells. [0003] With the development of surface chemistry and microfluidic technology, the controllable arrangement of different kinds of cells on the same substrate can precisely control the in vitro co-culture of cells, providing a new platform for cell biology, tissue engineering construction and drug scr...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12M3/04C12N5/00
Inventor 谢赟燕王黎明蒋兴宇
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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