Microflow cell culture array and application thereof

A cell culture and microfluidic technology, which is applied in the field of microfluidic technology and cell biology, can solve the problems of complex micromachining technology, inconvenient cultivation and observation of non-adherent cells, and the inability to observe the growth of cells for a long time. The effect of micro-observation convenience

Inactive Publication Date: 2008-10-01
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although these methods are useful, they require special surface treatment and require complex microfabrication processes or manipulations, making cell arrays in microfluidic systems not widely available to biologists.
Moreover, in these methods, the supply of nutrients to cells is mainly achieved through the flow of fluid around the cells, and the cells will be under a

Method used

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  • Microflow cell culture array and application thereof
  • Microflow cell culture array and application thereof
  • Microflow cell culture array and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Example 1 Microfluidic cell (microbe) culture array

[0028] Such as figure 1 As shown, the width of the main channel 1 is 100 microns, the cell culture chamber 2 is an equilateral triangle with a side length of 125 microns, and the apex of the triangle is connected to the main channel 1 through a diffusion channel 3 with a width of 20 microns and a length of 20 microns. The heights of the main channel 1 , the culture chamber 2 and the diffusion channel 3 are all 6 microns. The cell culture chamber 2 and the diffusion channel 3 constitute the cell culture unit 4, and several repeated cell culture units 4 are arranged on both sides of the main channel 1, constituting the basic unit of the microfluidic cell (microbe) culture array. The microfluidic cell (microbe) culture array of the present invention is composed of one or more microfluidic cell (microbe) culture array units. figure 2 Shown in is a microfluidic cell (microbe) culture array composed of 7 microfluidic ce...

Embodiment 2

[0031] Example 2 Observation of single cell culture

[0032] In this example, the microfluidic cell (microorganism) culture array ( figure 1 ) for single-cell culture observation of yeast cells.

[0033] specific method:

[0034] (1) Cultivate yeast cells in YPG culture medium, shake evenly before use and dilute to an appropriate concentration, the concentration is about 10 7 cells / ml.

[0035] (2) Take the chip and put it in a vacuum environment for about 10 minutes, inject the bacteria from the inlet, and the pushing speed is 20 microliters / hour. Such as image 3 , the yeast cells are automatically drawn into the cell culture unit ( image 3 a), when the cell culture unit is filled with solution ( image 3 b), the cell solution in the main channel can be washed away with the culture medium, and the yeast cells are cultivated, such as Figure 4 .

[0036] Studies have shown that through the above method, the yeast cell solution is automatically sucked into the cell cu...

Embodiment 3

[0037] Example 3 Observation of Effects of Environmental Changes on Cell Phenotypes

[0038] In this example, the microfluidic cell (microorganism) culture array ( figure 2 ) to observe and count the growth of yeast cells in different culture environments.

[0039] specific method:

[0040] (1) Culture yeast cells containing copper ion-induced green fluorescent protein in YPG culture medium, shake evenly before use and dilute to an appropriate concentration, the concentration is about 10 7 cells / ml.

[0041] (2) Take the chip and put it in a vacuum environment for about 10 minutes, inject the bacteria from the A inlet, and the pushing speed is 40 microliters / hour, so that the culture chambers in the seven cell culture channels are all filled with the bacteria solution.

[0042] (3) Channels 1 to 7 of inlet b are respectively fed with culture solutions containing different copper ion concentrations, and the copper ion concentrations are 0; 0.2; 0.4; 0.6; 1; 1.5; 2mM. Cell ...

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Abstract

The present invention provides a microorganism culture array, including one or more microflow cell culture array units, the microflow cell culture array unit includes a main channel and a plurality of cell culture units connected with the main channel, each culture unit includes a diffusion channel and a cell culture room, the cell culture room is connected with the main channel. The microorganism culture array of the invention not only realizes isolation of single-cell, but also makes the cell locate in zero flow state when the culture liquid or other reagent is changed, bringing greatly convenience for the cell culture and observation especially the culture and observation of non-adherent cells, greatly fit for research work of high throughput screen based on the cell phenotype.

Description

technical field [0001] The invention belongs to the field of microfluidic technology and cell biology, and in particular relates to a microfluidic chip device suitable for culturing cells or microorganisms in a microenvironment. Background technique [0002] Microfluidic chip (Microfluidic chip) originally originated in the field of analytical chemistry. It is a kind of micro-channel network structure and other functional units made on a substrate of several square centimeters by using fine processing technology to realize the preparation of micro-sample, processing A fast, efficient and low-consumption micro-analysis experimental device integrating sample, reaction, separation and detection. [0003] The application of microfluidic chip technology in cell culture and its biochemical analysis has attracted widespread attention, such as cell manipulation, expression of green fluorescent protein, gene transfection, cell activity testing, cell separation, measurement of intrace...

Claims

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Application Information

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IPC IPC(8): C12M1/00C12N1/14C12N1/16C12N1/20C12P1/02
Inventor 罗春雄欧阳颀
Owner PEKING UNIV
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