Microfluidic chip used for cell co-culture and cell culture method thereof

A microfluidic chip and cell culture technology, which is applied in the fields of biomedical engineering and cell biology research, can solve the problems of high reagent consumption, difficulty in high-throughput research, and inaccurate research results, and achieve simple processing and low cost. Shear force, the effect of simplifying the experimental procedure

Active Publication Date: 2017-03-29
BEIJING INSTITUTE OF TECHNOLOGYGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] However, the existing cell co-culture technology is mostly developed based on the traditional cell culture technology, and there are still some defects when applied to biological research.
First of all, most of the existing cell co-cultivation technologies are based on static cell culture, so the cultured cells are in a static microenvironment, while the cells in a multicellular organism are in a microenvironment jointly created by the circulatory s

Method used

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  • Microfluidic chip used for cell co-culture and cell culture method thereof
  • Microfluidic chip used for cell co-culture and cell culture method thereof
  • Microfluidic chip used for cell co-culture and cell culture method thereof

Examples

Experimental program
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Example Embodiment

[0041] Example 1

[0042] The invention provides a microfluidic chip for cell co-cultivation, such as figure 1 As shown, the microfluidic chip includes a base layer 1, a cell culture chamber layer, and an upper cover layer 4 from bottom to top. The cell culture chamber layer is formed by connecting several cell culture chambers on the same plane. For cell culture, Example 1 is an example of two cell culture chambers. The number of cell culture chambers of the present invention is not limited to two, and may be formed by multiple cell culture chambers communicating with each other.

[0043] The basal layer can be a transparent material processed by tissue culture according to the needs of the type of cultured cells; or a transparent material that meets the requirements of cell culture through coating: such as polyethylene terephthalate PET, glass or polystyrene PS Wait.

[0044] The cell culture chamber layer is provided with a screen 22 to divide each cell culture chamber into two u...

Example Embodiment

[0049] Example 2

[0050] Preferably, an observation area is provided on the upper cover layer and the cell culture chamber layer corresponding to adjacent positions of the two cell culture chambers, such as figure 1 As shown, the position of the observation area is not limited in the present invention, and all areas that can realize the observation of cells in two adjacent cell culture chambers are regarded as observation areas. The observation area can use any transparent non-biologically toxic material as the upper cover plate to observe and detect the cells in two adjacent cell culture chambers.

Example Embodiment

[0051] Example 3

[0052] An intermittent culture method for cell co-culture of human neuroblastoma SH-SY5Y and human glioma U87MG on a microfluidic chip cultured cells. In this embodiment, the structure of the microfluidic chip may not be provided with a filter Net and sieve silk make the microscopic image clear.

[0053] The method includes the following steps:

[0054] Human neuroblastoma SH-SY5Y and human glioma U87MG were frozen and stored at -150℃. Before the experiment, the cells were recovered and placed at 37℃, 5% CO 2 In an incubator, culture in DMEM medium, and add 10% fetal bovine serum, 100 U / mL penicillin G, 100 μg / mL streptomycin, and 2% MEM NEAA (100X) to the medium. The cells to be conventionally cultured can grow to the logarithmic phase and then be seeded into the microfluidic chip.

[0055] Take the trypsin-digested cells, and terminate the digestion with fetal bovine serum after the digestion. The digested cells were centrifuged at 1000 rpm for 5 minutes, and th...

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Abstract

Disclosed is a microfluidic chip used for cell co-culture and a cell culture method thereof. The microfluidic chip comprises successively from bottom to top: a basal layer, a cell culture chamber layer and an upper cover plate layer. A bolting cloth is arranged in the cell culture chamber layer to divide cell culture chambers into two layers, the upper layer is a culture medium exchanging layer, and the lower layer is a cell culture layer. A microfiltration membrane is arranged between the adjacent cell culture chambers. One or more upper cover plate through holes, corresponding to each cell culture chamber, are formed in the upper cover plate layer, and the through holes mutually correspond to bolting cloth through holes in position. Connectors are arranged in the positions corresponding to the upper cover plate through holes. A pipeline used for culture medium exchanging and cell inoculating or cell detecting is arranged in each connector. The microfluidic chip used for cell co-culture can intercept and retain suspension cells effectively, so that microfluidic cell co-culture of adherent cells and suspension cells or suspension cells and adherent cells is achieved.

Description

technical field [0001] The invention belongs to the field of biomedical engineering and cell biology research, and in particular relates to a microfluidic chip for co-culture of cells and a cell culture method thereof. Background technique [0002] Cell culture is a basic experimental method for various research in life sciences. Although the development of this technology has slowed down in recent years, cell culture has still made great contributions to biological research in many fields for a long time. However, with the deepening and expansion of research, many studies have more and more new requirements for cell culture, such as the need to maintain cell activity in a microvolume environment for a long time, effective in microgravity and other special environmental conditions. cell culture etc. Therefore, people began to explore and develop new cell culture techniques. [0003] At the same time, due to the huge difference between the environment of cells in single cel...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M3/06C12N5/09
CPCC12M23/16C12M23/22C12M23/34C12M29/10C12M33/14C12M35/08C12N5/0693C12N5/0694C12N2502/30C12N2533/52
Inventor 马宏陈钰王品虹邓玉林于世永李瑞
Owner BEIJING INSTITUTE OF TECHNOLOGYGY
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