JoVE Encyclopedie van Experimenten
Kankeronderzoek
0 weergaven • 3:14 min. • April 30th, 2023
- Pancreatic cancer stem cells or CSCs are a subset of self-renewable cancer cells and can give rise to the entire tumor. CSCs reside in a well-protected niche consisting of other cells, such as fibroblasts, immune cells, endothelial cells, and extracellular matrix components, such as cytokines and growth factors. The interactions happening between the cells in such an environment help CSCs propagate and increase tumor growth.
To study the properties of CSCs, we isolate them from the pancreatic tumors. Begin by taking the cell suspension from human pancreatic tumor tissue in a CSC growth media. Next, seed the suspension, consisting primarily of fibroblasts and CSCs, on a gelatin-coated plate and incubate it for the desired duration. Fibroblasts adhere to the gelatin surface, leaving the free-floating CSCs in the cell suspension.
Take a small quantity of cell suspension and add trypan blue. Pipette this mixture to a hemocytometer and count the number of viable cells. Trypan blue enters the dead cells and stains them blue leaving the live cells colorless. Finally, store the CSCs for further analysis.
In the following protocol, we will isolate the CSCs from pancreatic tumors.
- To iso
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