The cell infiltration growth assay test can be applied to (1) detect cancer cell traits and (2) study the mode of cell infiltration or penetration.

Operation method

Cell Infiltration Growth Assay

Principle

Infiltrative growth is the ability of cells to infiltrate or penetrate other tissues to proliferate and grow, and infiltrative growth test is a meaningful indicator for detecting cancerous cells. The test is performed by co-culturing other normal tissues with cancerous cells and observing the infiltration and growth of cancerous cells into normal tissues.

Materials and Instruments

Chicken embryo
Bacto Agar Eagle Buffer Calf Serum Formalin Hanks Liquid Iodine
Glass Rings CO2 Warmers Petri dishes Flat dish lids Surgical scissors Surgical forceps Centrifuge Syringes Beakers

Move

Chicken embryo skin or myocardial tissue mass is commonly used as the object to be infiltrated and grown; now take chicken embryo skin as an example, the culture process is as follows
1. Procedure
(1) Preparation of chicken embryo leachate
(2) Agar layer preparation
1 % Bacto agar (prepared in NaHCO3-free Eagle's buffer) 10 pts.
Calf serum 4 points
Chicken embryo extract 4 marks
(3) Chicken embryo skin culture

Aseptically remove the skin from the back of 10-day-old chicken embryo, cut into ^6-8mm3 pieces; place 2-3 pieces in each dish and stick them on the surface of agar layer;
(4) Take a sterile small magnetic cup or glass ring (silicone rubber ring can also be used) with an inner diameter of 6 mm and a thickness of 2 mm, and put it on the small pieces of chicken skin;
(5) Co-culture

(5) add 25 ~ 30 ul cell suspension (containing ¹⁰⁴ cells of 2 times the culture solution) to each small magnetic circle, cover the dish with a lid, and place it in a 37 ℃ _CO2 incubator for culture;
(6) Replenishment

On the next day, add 2.5 ul of 2-fold culture medium to each ring;
(7) Slice observation

On the 4th day, excavate and take out the chicken skin block, fix it with 10% formalin, embed it routinely, section it, stain it, and observe the infiltration of the cells into the skin of chicken embryo under the light microscope.
2. Results

If the cancerous cells have infiltrated into the chicken embryo skin, it means that the cells have the ability to infiltrate and grow.

Caveat

Bacterial infection should be avoided during the experiment. If the cancerous cells have infiltrated into the skin of the chicken embryo, it means that the cells have the ability to infiltrate and grow.

Common Problems

Chicken embryo leachate contains growth factors, large nuclear proteins and small molecules of amino acids, etc., which have the effect of stimulating cell growth, and it is the culture fluid used in the early years of culture, which has now been replaced by the synthetic culture fluid, but it still has some application value in certain research.

Source "Tissue culture and molecular cytology technology".

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