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Method for culturing cells under pressure condition

A technology for culturing cells and cells, which is applied in tissue culture, biochemical equipment and methods, and specific-purpose bioreactors/fermenters, etc. It can solve problems such as poor repeatability, different gel sizes, and poor gel formation effects, etc. problem, to achieve stable pH value and good gel quality

Inactive Publication Date: 2022-04-15
THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0003] The inventors found that the existing method has poor gel formation effect, the gel compounding liquid is easy to leak from the sponge ring, the repeatability is poor, the amount of cells carried by the gel is small, and the size of the gel is different, so it is impossible to ensure that each piece of gel can be fully absorbed during pressure culture. Glue under the same pressure and other defects

Method used

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  • Method for culturing cells under pressure condition
  • Method for culturing cells under pressure condition
  • Method for culturing cells under pressure condition

Examples

Experimental program
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Effect test

Embodiment 1

[0040] 1. Preparation of Rat Tail Type I Collagen 3D Culture Gel and Pressurized Culture

[0041] 1) Take a 24-well plate, each well as a culture unit, and add in the following order: ① 42 μl 10*PBS; ② 18 μl NaOH (0.1mol / L); ③ 300 μl mouse tail type 1 collagen (Hangzhou Xinyou Biotechnology Company, 5 mg / ml) ; ④ 1ml cell suspension, the cell suspension was resuspended human dermal microvascular endothelial cells (different from sponge ring cells) in F12 DMEM containing 10% FBS. After adding the sample, pipette 5 times to mix quickly. 37°C, 5% CO 2 Let the incubator stand for 10 minutes to form a gel.

[0042] The resulting gel had a pH of about 7.3 and was translucent and light orange, and the cells were observed to be stretched under an optical microscope. The gel is molded in advance with the help of the shape of each well of the 24-well plate, which can ensure that the height of each gel block is consistent after formation. In order to verify the stability of the gel, a...

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Abstract

The invention relates to a method for culturing cells under a pressure condition, which comprises the following steps of: pre-forming gel for carrying the cells, transferring the cells together with the gel into a sponge ring of a pressure culture plate, and culturing the cells under pressure. According to the method, the gel for bearing the cells is formed in advance, the formation of the gel is not influenced by the sponge ring, the high-quality gel can be formed, the pressure borne by the cells in the gel is the same, and the growth of the cells is not influenced by the sponge ring.

Description

technical field [0001] The invention relates to the fields of mechanical force conduction and mechanobiological effect culture, in particular to a method for culturing cells under pressure conditions. Background technique [0002] At present, research on the influence of mechanobiological effects on cells is widely carried out. It has become a research hotspot to use pressure culture systems (such as Flexcell pressure culture system) to simulate changes in the mechanical microenvironment of the human body, and to explore the mechanobiological effects of mechanical pressure and the mechanism by which cells feel mechanical pressure. The current common pressure culture method is to use the Flexcell cell mechanical loading system to directly load the prepared cell-containing gel liquid into the sponge ring of the pressure culture plate, and rely on the surface tension of the liquid to form a gel in the sponge ring, and then add pressure culture. [0003] The inventors found tha...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C12M1/00
Inventor 李猛智舒斌祁少海
Owner THE FIRST AFFILIATED HOSPITAL OF SUN YAT SEN UNIV