This video describes the culturing of breast cancer cells adjacent to femur bone explants to measure cancer cell proliferation and colonization in a human bone tissue explant model system.
Protocol
1. Co-culturing Breast Cancer Cells Adjacent to Bone Fragments to Measure Breast Cell Proliferation Using BLI Prior to the experiment, prepare a suspension containing 2 x 105 breast cancer cells/50 µl, and prepare plugs of bone wax for immobilizing bone fragments in culture. Cut a P200 micropipette tip into 3 pieces and use the narrow end of the largest piece in a “cookie cutter” manner to cut small (~35 mg) pieces of bone wax. Use the wide end of the smallest piece to eject t…
Divulgazioni
The authors have nothing to disclose.
Materials
DMEM (1X) + GlutaMAX-l
Life Technologies
10569-010
Supplement all DMEM with 10% fetal bovine serum and 1% Pen/ Strep
Fetal bovine serum
Life Technologies
16000-044
Penicillen Streptomycin
Life Technologies
15140-122
D-luciferin firefly, potassium salt 5 g (30 mg/ml)
Co-culturing BC Cells: Culturing Breast Cancer Cells with Bone Fragments to Study Cancer Cell Proliferation. J. Vis. Exp. (Pending Publication), e20215, doi: (2023).