Monitoring Dynamic Interactions Between Breast Cancer Cells and Human Bone Tissue in a Co-culture Model

Abstract

Purpose Bone is a preferential site of breast cancer metastasis and models are needed to study this process at the level of the microenvironment. We have used bioluminescence imaging (BLI) and multiplex biomarker immunoassays to monitor dynamic breast cancer cell behaviors in co-culture with human bone tissue. Procedures Femur tissue fragments harvested from hip replacement surgeries were co-cultured with luciferase-positive MDA-MB-231-fLuc cells. BLI was performed to quantify breast cell division and track migration relative to bone tissue. Breast cell colonization of bone tissues was assessed with immunohistochemistry. Biomarkers in co-culture supernatants were profiled with MILLIPLEX® immunoassays. Results BLI demonstrated increased MDA-MB-231-fLuc proliferation (p<0.001) in the presence vs. absence of bones, and revealed breast cell migration toward bone. Immunohistochemistry illustrated MDA-MB-231-fLuc colonization of bone, and MILLIPLEX® profiles of culture supernatants suggested breast/bone crosstalk. Conclusions Breast cell behaviors that facilitate metastasis occur reproducibly in human bone tissue co-cultures and can be monitored and quantified using BLI and multiplex immunoassays.

An in vitro 3D bone metastasis model by using a human bone tissue culture and human sex-related cancer cells

“…Unlike other reports [18, 25] we did not observe a decline in bone marrow viability after 7 days (T1) of culture as revealed by the Alamar Blue test and appearance in histological sections. In fact, the bone viability results obtained immediately after surgery (T0) were maintained after 7 days (T1) of culture with and without cancer cells (MCF-7 and PC-3), in both normoxic and hypoxic conditions.…”

“…Currently, with the growing acceptance of in vitro models as effective tools for studying cancer biology, many enhanced and novel 3D models have emerged in an effort to recapitulate the native tumor and its microenvironment [17–25]. We developed a 3D model of bone metastases using human bone tissue taken from total hip replacement surgery patients cultured with human breast or prostate cancer cells (MCF-7 and PC-3) and established a “proof of concept” to recapitulate the bone metastasis and its microenvironment.…”

“…age, sex, concomitant comorbidity and unhealthy life style factors. Recently, Contag et al [25] developed a co-culture model able to monitor dynamic interactions between human breast cancer cells and human bone tissue. The authors used a static model where the bone marrow was depleted from the bone tissue and cancer cell/bone interactions were evaluated during relatively short time periods (for up to 96 hours) and not in hypoxic conditions, which is typical of tumors grown in vivo .…”

Prevention of breast cancer skeletal metastases with parathyroid hormone

“…Murine 4T1 and human MDA-MB-231 breast cancer cells obtained originally from ATCC were genetically engineered to stably express firefly luciferase and enhanced green fluorescence protein using the Sleeping beauty transposon plasmid pKT2/LuBiG (72,73). These cells are referred to as MDA (a gift from Bonnie King, Stanford University) and 4T1 (a gift from Michael Bachmann, Stanford University).…”

Methods for Culturing Human Femur Tissue Explants to Study Breast Cancer Cell Colonization of the Metastatic Niche

“…Incubate the receiver plate in the tissue culture incubator while seeding the inserts. Note: The remaining supernatant (~1 ml after evaporation) can be collected for analysis of secretome factors if desired 44 . 3.…”