JoVE Encyclopedia of Experiments
Cancer Research
0 views • 3:01 min • April 30th, 2023
Certain organs produce soluble factors and become preferential sites for breast cancer metastasis. A conditioned medium prepared from these harvested organs cells may help identify and study such factors.
Begin by placing a harvested organ such as a liver in a tube containing cold PBS and flipping the tube up and down to remove the organ's residual blood. Place the organ in a Petri dish. And using a scalpel, dice the organ into small pieces.
Resuspend these pieces into a tube containing a basal medium supplemented with an antibiotic to support cell growth and prevent contamination. Now, pour the cell suspension into a culture plate and incubate at 37 degrees Celsius with a continuous carbon dioxide supply for the required duration.
During incubation, these cells release metabolites, growth factors, extracellular matrix proteins into the medium. Next, transfer the cell suspension to a centrifuge tube and add fresh medium to dilute it. Centrifuge the tube and filter the supernatant, which is the conditioned medium, into a fresh tube.
In the following protocol, we will prepare conditioned media from the mouse lungs, and brain to study the metastatic behavior of breast cancer cells.
To
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