title: Methods for culturing human femur tissue explants to study breast cancer cell colonization of the metastatic niche.
abstract: Bone is the most common site of breast cancer metastasis. Although it is widely accepted that the microenvironment influences cancer cell behavior, little is known about breast cancer cell properties and behaviors within the native microenvironment of human bone tissue.We have developed approaches to track, quantify and modulate human breast cancer cells within the microenvironment of cultured human bone tissue fragments isolated from discarded femoral heads following total hip replacement surgeries. Using breast cancer cells engineered for luciferase and enhanced green fluorescent protein (EGFP) expression, we are able to reproducibly quantitate migration and proliferation patterns using bioluminescence imaging (BLI), track cell interactions within the bone fragments using fluorescence microscopy, and evaluate breast cells after colonization with flow cytometry. The key advantages of this model include: 1) a native, architecturally intact tissue microenvironment that includes relevant human cell types, and 2) direct access to the microenvironment, which facilitates rapid quantitative and qualitative monitoring and perturbation of breast and bone cell properties, behaviors and interactions. A primary limitation, at present, is the finite viability of the tissue fragments, which confines the window of study to short-term culture. Applications of the model system include studying the basic biology of breast cancer and other bone-seeking malignancies within the metastatic niche, and developing therapeutic strategies to effectively target breast cancer cells in bone tissues.
authors: Department of Pediatrics, Stanford University School of Medicine.
authors: Zachary S
authors: ZS
authors: Templeton
authors: Department of Pediatrics, Stanford University School of Medicine.
authors: Michael H
authors: MH
authors: Bachmann
authors: Department of Pediatrics, Stanford University School of Medicine.
authors: Rajiv V
authors: RV
authors: Alluri
authors: Department of Orthopaedic Surgery, Stanford University School of Medicine.
authors: William J
authors: WJ
authors: Maloney
authors: Department of Pediatrics, Stanford University School of Medicine.
authors: Christopher H
authors: CH
authors: Contag
authors: Department of Pediatrics, Stanford University School of Medicine; bonnie.king@stanford.edu.
authors: Bonnie L
authors: BL
authors: King
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001859:Bone Neoplasms / Q000473:pathology / Q000556:secondary*; D001943:Breast Neoplasms / Q000473:pathology*; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000502:physiology; D005260:Female; D005269:Femur / Q000166:cytology* / Q000473:pathology; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D008875:Middle Aged; D009362:Neoplasm Metastasis; D014411:Neoplastic Stem Cells / Q000473:pathology; D055153:Stem Cell Niche; D046509:Tissue Culture Techniques / Q000379:methods*; D059016:Tumor Microenvironment