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'Funnel'-shaped three-dimensional cell aggregation and culture chip and preparation method thereof

A three-dimensional cell and funnel technology, used in tissue cell/virus culture devices, biochemical equipment and methods, and methods for supporting/immobilizing microorganisms, etc.

Pending Publication Date: 2018-11-13
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, at present, most of the chips with concave structures prepared by soft lithography have vertical concave sidewalls, which are difficult to promote cell aggregation into clusters or balls to a certain extent.

Method used

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  • 'Funnel'-shaped three-dimensional cell aggregation and culture chip and preparation method thereof
  • 'Funnel'-shaped three-dimensional cell aggregation and culture chip and preparation method thereof
  • 'Funnel'-shaped three-dimensional cell aggregation and culture chip and preparation method thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0027] A method for preparing a "funnel"-like three-dimensional cell aggregation culture chip, characterized in that it is carried out in accordance with the following steps:

[0028] (1) Spin-coat SU-8 photoresist with a thickness of 400 microns on the surface of the glass substrate, and bake it at 95°C for 4 hours,

[0029] (2) Tilt angle exposure: fix a square array mask with a side length of 350 microns on the surface of the substrate containing photoresist, and place it on a platform that can adjust the tilt angle and rotate freely.

[0030] (3) During UV exposure, expose 4 times along the apex with an inclination angle of 60 degrees, and expose 4 times along the edge with an inclination angle of 40 degrees and rotate 45 degrees each time.

[0031] (4) Post-bake at 95°C for 10-30 minutes, add 400 micron thick SU-8 photoresist, and bake for 4 hours at 95°C for the second time.

[0032] (5) Vertical exposure: Use the same square mask with a side length of 350 microns to ex...

Embodiment 2

[0036] A method for preparing a "funnel"-like three-dimensional cell aggregation culture chip, characterized in that it is carried out in accordance with the following steps:

[0037] (1) Spin-coat SU-8 photoresist with a thickness of 300 microns on the surface of the glass substrate, and bake it at 95°C for 4 hours,

[0038] (2) Tilt angle exposure: fix a square array mask with a side length of 350 microns on the surface of the substrate containing photoresist, and place it on a platform that can adjust the tilt angle and rotate freely.

[0039] (3) During ultraviolet exposure, expose 4 times along the apex with a tilt angle of 50 degrees, and expose 4 times along the edge with a tilt angle of 35 degrees and rotate 45 degrees each time.

[0040] (4) Post-bake at 95°C for 10-30 minutes, add 500 micron thick SU-8 photoresist, and bake for 4 hours at 95°C for the second time.

[0041] (5) Vertical exposure: After the circular array mask of a square with a side length of 350 mic...

Embodiment 3

[0045] A method for preparing a "funnel"-like three-dimensional cell aggregation culture chip, characterized in that it is carried out in accordance with the following steps:

[0046] (1) Spin-coat SU-8 photoresist with a thickness of 300 microns on the surface of glass or silicon substrate, and bake it at 95°C for 4 hours,

[0047] (2) Tilt angle exposure: Fix a square array mask with a side length of 400 microns on the surface of the substrate containing photoresist, and place it on a platform that can adjust the tilt angle and rotate freely.

[0048] (3) During ultraviolet exposure, expose 4 times along the apex with a tilt angle of 50 degrees, and expose 4 times along the edge with a tilt angle of 35 degrees and rotate 45 degrees each time.

[0049] (4) Post-bake at 95°C for 10 minutes, add a 450-micron thick SU-8 photoresist, and bake for 8 hours at 95°C for the second time.

[0050](5) Vertical exposure: After the circular array mask of a square with a side length of 40...

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Abstract

The invention provides a 'funnel'-shaped three-dimensional cell aggregation and culture chip and a preparation method thereof. The chip is provided with a 'funnel'-shaped recessed array structure, andthe upper half portion of the funnel-shaped structure is a trapezoidal pyramid while the lower half portion of the funnel-shaped structure is a cylinder or a cuboid. According to a preparation process of the chip, a template which has a funnel-shaped structure is obtained by two soft lithography technologies including oblique exposure and vertical exposure at first, then a polydimethylsiloxane chip is prepared by the template, and when ultraviolet exposure is carried out by soft lithography, the upper half portion and the lower half portion of the funnel-shaped structure are separately prepared by two exposure methods. After the chip is inoculated with cells, because an inclined side wall is arranged on the top of a funnel-shaped recess in the chip, cells are forced to be aggregated to the bottom of the funnel-shaped recess and then are aggregated into cell groups or cell balls.

Description

technical field [0001] The invention belongs to the micro-manufacturing process of material science, and in particular relates to the preparation of a "funnel"-like three-dimensional cell aggregation culture chip by using a soft photolithography method and a preparation method thereof. Background technique [0002] Traditional 2D monolayer cell culture cannot truly reflect the living environment of cells in vivo, which has become an insurmountable gap for in vitro cell experiments to simulate physiological states. In contrast, cell 3D culture technology can narrow the gap between in vitro experiments and in vivo experiments, and better reflect the idea of ​​bionics. Cell sphere culture is a kind of 3D culture. In addition, cells aggregated into spheres can form embryoid bodies, which is convenient for subsequent induction and differentiation, and can be used for in vitro pathology, toxicology research and drug screening. [0003] At present, the existing cell sphere culture...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00G03F7/00
CPCG03F7/0035C12M25/14
Inventor 秦建华魏文博刘海涛
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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