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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 system and a variety of cells. In the dynamic microenvironment of , this difference may lead to inaccurate research results
Second, the existing cell co-culture technology is difficult to achieve high-throughput research
Finally, the reagent consumption of the existing cell co-culture technology is relatively large, especially for the research of some precious samples, researchers often cannot directly use the existing cell co-culture technology for research

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

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Embodiment 1

[0042] The invention provides a microfluidic chip for co-cultivation of cells, such as figure 1 As shown, the microfluidic chip includes a base layer 1, a cell culture chamber layer, 2, and an upper cover layer 4 from bottom to top. The cell culture chamber layer is composed of several cell culture chambers connected to each other on the same plane. For cell culture, Example 1 is an illustration of two cell culture chambers. The number of cell culture chambers in the present invention is not limited to 2, and can be formed by connecting multiple cell culture chambers.

[0043] The base layer can be a transparent material that has been treated with tissue culture according to the needs of the cultured cell type; 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 sieve 22 to divide each of the cell culture chambers ...

Embodiment 2

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

Embodiment 3

[0052] A microfluidic chip culture method for intermittent culture of cells co-cultivating human neuroblastoma SH-SY5Y and human glioma U87MG. In this embodiment, the structure of the microfluidic chip may not be provided with a filter Mesh and sieve to make microscopic imaging clear.

[0053] The method includes the following steps:

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

[0055] The cells were digested with trypsin, and the digestion was terminated with fetal bovine serum after digestion. The digested cells were centrifuged at 1000rpm for 5min, and the culture medium was u...

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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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