Co-culturing BC Cells: Culturing Breast Cancer Cells with Bone Fragments to Study Cancer Cell Proliferation

Start by placing a piece of bone wax into two cell culture plates labeled as test and control. Pipette an appropriate amount of bioluminescence expressing breast cancer cells at each plate center and incubate the plates for a specific duration to aid cell attachment. Place a small bone explant on the bone wax in the test plate and secure it by applying pressure.

Slowly pour a cell culture medium with fetal bovine serum, a growth supplement, to each plate side to submerge the explant and incubate at 37 degrees Celsius for a specific duration. The bone tissue in the test plate releases soluble factors, such as cytokines and SRC kinase, which attract breast cancer cells, resulting in their migration. Add a substrate to each plate and place the plates on a bioluminescence imaging chamber.

The cancer cell expressing bioluminescent protein oxidizes the substrate and emits light. Quantify proliferation by measuring total photons emitted by each cell using the software. Cancer cells in the test plate should proliferate faster than the cells in the control plate. In the following protocol, we will co-culture breast cancer cells adjacent to femur bone explants to measure breast cell prolifer