Authors
Johannes C. Reichert, Les J. Burke, Judith A. Clements, and Dietmar W. Hutmacher
Summary
Suitable matrices to study metastasis and cancer cell growth in the bone microenvironment have been limited to matrices consisting of specific proteins such as collagen or those secreted by immortalised cell lines. Use of the latter is restrictive since these lines retain characteristics representative of a particular stage of differentiation and often do not have the capacity to differentiate further. Thus, the matrix generated by the immortalised cell secretome will not be representative of the normal bone environment first seen by a tumour cell in the metastatic process. We describe a method to produce and analyse matrices secreted by human primary osteoblasts grown in medium which promotes secretion of a mineralised matrix. Osteoblasts can be readily and efficiently removed to leave matrices which are free of cells, show a high degree of calcification and contain collagen type I which is representative of the bone matrix in vivo. These matrices are routinely used for culture of two prostate cancer cells lines, PC3 and LNCaP, which are easily harvested for molecular analyses. This in vitro system offers a more physiologically relevant environment to study the interaction of tumour cells and the bone microenvironment.Further details
The protocol was published on Protocol Exchange in 2008. To see the entire protocol, click on the source link.Advertisement
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