Introduction to cell scratch test

Cell Scratch test (Wound Healing) is a simple, affordable Test Method of in vitro cell migration / invasion of the tumor. The principle of this method is that when the cells grow to merge into a single layer state, a blank area is artificially created on the fused monolayer cells, called "scratches." The cells at the edge of the scratch will gradually enter the blank area to cause the "scratches" to heal.

The basic steps include the manufacture of "scratches", the acquisition of images during cell migration, and the processing of later data.

Features:

1, to some extent simulate the process of cell migration in vivo.

2, very suitable for studying cell migration with extracellular matrix (ECM), cell-to-cell interaction.

3. Compatible with microscope systems including live cell imaging and can be used to analyze interactions between cells.

4. The easiest way to study cell migration in vitro.

Traditional experimental method

Experimental steps:

1. Before inoculation of the cells, use a marker pen to draw horizontal lines on the back of the 12-well plate (to locate the same field of view when taking pictures).

2, after the cells are digested, access to the 12-well plate, the number is to cover the bottom of the plate after the wall is suitable (if the number is small, it can be cultivated for a period of time to cover the bottom of the plate).

3. After the cells are covered with the bottom of the plate, use a 1ml tip to make cell scratches perpendicular to the orifice plate, and try to ensure that the width of each scratch is consistent. (The scratches made by the artificial gun head are difficult to ensure the consistency of the scratch width and affect the experimental results, which is also the biggest flaw of the method.)

4. Aspirate the cell culture medium and rinse the plate three times with PBS to wash away the cell debris produced by the scratches.

5. Add serum-free medium and take a photo record.

6. The culture plate is placed in an incubator and taken out every 4-6 hours.

7. Analyze the experimental results based on the collected image data.

German IBIDI company Culture Insert method

1, prepare cells, culture, culture Insert.

2, as shown in the figure, the cells were inoculated into the Insert in the middle of Dish. After the cells were overgrown in the Insert area, the Inserts were removed with tweezers to produce scratches of 500 μm width.

3, take pictures every 4-6 hours.

4. Analyze the experimental results based on the collected image data.

Experimental principle diagram

Summary: Compared to traditional scratch experiments, Ibidi's design is more scientific and reproducible.

references:
A Novel Sialyltransferase Inhibitor Suppresses FAK/Paxillin Signaling and Cancer Angiogenesis and Metastasis Pathways
JY Chen, YA Tang, SM Huang, HF Juan, LW Wu, YC Sun, SC Wang, KW Wu, G. Balraj and TT Chang
Cancer Research , 2011, 10.1158/0008-5472.CAN-10-1303

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