Cell invasion chemotaxis model and preparation method thereof

A cell and model technology, applied in animal cells, tumor/cancer cells, vertebrate cells, etc., can solve the problem of not excluding gravity factors

Inactive Publication Date: 2016-04-27
SOUTHERN MEDICAL UNIVERSITY
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Problems solved by technology

[0003] Although the Transwell experiment can prove the phenomenon of cell invasion, chemotaxis and migration, there are the following problems: (1) it observes the cell movement indirectly; (2) it does not exclude the factor of gravity; (3) it cannot detect the expression of related proteins

Method used

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  • Cell invasion chemotaxis model and preparation method thereof
  • Cell invasion chemotaxis model and preparation method thereof
  • Cell invasion chemotaxis model and preparation method thereof

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Experimental program
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Embodiment 1

[0034] 1: Model making (see the specific model figure 1 )

[0035] 1.1 Cell culture: culture colorectal cancer cells sw480, digest when the cells grow to 90%, make cell suspension, and then absorb 5x10 cells by cell counting 5 The cell suspension of the cells is ready for use (in actual operation, according to the needs of individual experiments in 5x10 5 -1x10 6 within this range to determine the number of cells);

[0036] 1.2 Matrigel configuration of the lower layer: The selected consumables and reagents are as follows: 0.2ml transparent cap PCR thin-walled tube purchased from Axygen; Matrigel from BD Company; 1640 culture medium from hyclone; fetal bovine serum from hyclone. First, matrigel and tumor cell culture medium (hyclone's 1640 culture medium) are configured at a volume ratio of 37.8 μl: 25.2 μl (configured according to personal experiment requirements, preferably matrigel and hyclone's 1640 culture medium at a volume ratio of 3: 2-1:1), then add 7 μl of serum ...

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Abstract

The invention discloses a cells invasion and chemotaxis model as well as a preparation method thereof, which comprises the following steps: 2)culturing cells; 2)configuring a lower layer matrix glue; 3) configuring maker 1; 4)configuring an intermediate cell layer; 5) configuring maker 2; and configuring an upper layer matrix glue. The cells invasion and chemotaxis model can visually obtain the powerful evidence for cells chemotaxis and migration based on overcoming the gravity, and can perform HE coloration, imunohistochemistry and immunofluorescence, so that cell associated protein expression condition can be observed.

Description

technical field [0001] The invention relates to the field of cell invasion and chemotaxis, in particular to a cell invasion and chemotaxis model and a preparation method thereof. Background technique [0002] So far, the most commonly used experimental technology for tumor cell chemotaxis, migration, and invasion experiments is Transwell. The main material of this technology is Transwell chamber (Transwell chamber, Transwell insert), which is shaped like a small chamber that can be placed in a well plate. Cup, the bottom of the cup is a permeable membrane, and the material of the rest of the cup is the same as the ordinary orifice plate. This layer of membrane has micropores with a pore size of 0.1-12.0um. Different materials can be used according to different needs. Generally, polycarbonate membrane (polycarbonate membrane) is commonly used. When the results are counted, the cells are fixed and stained after the cells are taken out. Just count. [0003] Although the Trans...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09
Inventor 邓永键丁彦青马丽丽
Owner SOUTHERN MEDICAL UNIVERSITY
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