Interactions between breast cancer cells and bone marrow derived cells in vitro define a role for osteopontin in affecting breast cancer cell migration

Abstract

The preferential metastasis of breast cancer cells to bone is a complex set of events including homing and preferential growth which may include unique factors produced by bone cells in the immediate microenvironment. In this study, we evaluated the suitability of bone cells derived from orthoplastic surgeries for use in an in vitro co-culture system representing a model of the bone microenvironment. Using a limiting dilution assay we determined the relative survival and proliferation potentials of breast cancer cell lines co-cultured on bone-derived cells or on Hs68 fibroblasts. The comparison of bone and skin fibroblastic substrata indicates that MCF-7 cells preferentially survive and grow in a bone microenvironment (P < 0.001). Overall, we show that bone-derived cells enhance survival, proliferation, and migration of breast cancer cells, where migration is in part mediated by bone cell-produced osteopontin. Our in vitro co-culture model system provides a robust cost-effective method to study the various factors that mediate cancer/bone-derived cell interactions.

Role of Src in breast cancer cell migration and invasion in a breast cell/bone-derived cell microenvironment

“…Bone-derived cells were passaged in T75 flasks using Enzyme Cell Detachment Medium (Accutase, eBioscience, Cat # 00-4555-56) and cultured in 10% (v/v) FCS-DMEM with media changes twice weekly. These were shown previously to have mesenchymal stem cell characteristics [22]. Bone-derived cell cultures for all experiments were used between passage one and passage six.…”

“…We have previously developed an in vitro co-culture system comprised of bone mesenchymal stem cells (MSCs) cultured with breast cancer cell lines to study the factors mediating breast cancer cell migration and invasion in the breast/bone microenvironment [22]. This system allows the study of specific changes in gene expression in breast or bone-derived cells when cultured together and to test new therapeutics directed at preventing or treating breast cancer metastasis to bone.…”

“…This system allows the study of specific changes in gene expression in breast or bone-derived cells when cultured together and to test new therapeutics directed at preventing or treating breast cancer metastasis to bone. Using this model system, it has been previously shown that the bone sialoprotein osteopontin, when produced by bone MSCs, plays a significant role in breast cancer cell migration in the breast/bone microenvironment [22]. The current study investigates the role of Src tyrosine kinase on breast cancer cell migration and invasion in the breast or bone environment.…”

Breast Cancer Metastasis: Advances Through the Use of In Vitro Co-Culture Model Systems

“…There are a number of reports in the literature suggesting Osteopontin produced by breast cancer cells regulates their migratory properties and contributes to the aggressiveness of the disease (Sharp, Sung et al 1999;Chakraborty, Jain et al 2008;Hedley, Welch et al 2008;Patani, Jouhra et al 2008;Ribeiro-Silva and Oliveira da Costa 2008). In our co-culture model we found, at least in the breast/bone microenvironment, that it was Osteopontin produced by the bone cells, not the breast cells that increased breast cancer cell migration (Koro, Parkin et al 2010). …”

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

“…In vitro approaches for studying breast cancer cells in human bone co-cultures have typically focused on specific bone cell types such as marrow-derived osteoblasts or stromal cells cultured as monolayers or within engineered 3-dimensional model systems [25][26][27][28][29][30][31][32][33][34][35] . Although 2-dimensional cell culture models have formed the mainstay of in vitro approaches for cancer research, it has long been recognized that cell behavior is fundamentally altered in monolayer culture systems 18,36,37 .…”