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NC3Rs: National Centre for the Replacement Refinement & Reduction of Animals in Research
Pilot study grant

Development of human ex vivo bone-tumour niche for the study of cancer bone disease

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At a glance

Award date
November 2014 - September 2015
Grant amount
Principal investigator
Dr James Edwards
University of Oxford


  • Replacement
Read the abstract
View the grant profile on GtR

Application abstract

This proposal will standardise and validate the use of a human ex vivo assay for the study of cancer-bone disease, thereby replacing mice as the first stage model for the study of cancer cell interactions within the skeleton, and providing a new system to assess novel drugs aimed at reducing cancer metastasis to and proliferation within bone, in addition to blocking tumour-induced osteolysis.
Uniform slices (0.5cm thick) of human bone from cores (0.5cm diameter, approx. 5 cm length) isolated from femoral heads removed during surgery will be cultured ex vivo. Human tumour cells (breast cancer, myeloma) will be seeded within each bone slice at a known density using either A) direct injection over 30 mins, followed by an overnight colonisation and transfer to experimental well plate; B) implantation as above followed by 1hr gentle agitation prior to overnight colonisation; C) immersion of bone slice in matrigel and implantation of tumour cells within bone/gel environment, followed by culture as in A. All cultures will be maintained for either 3, 7 or 10 days in aMEM (+1%BSA) as used previously and replenished every 3 days. Anti-cancer therapies previously tested within our group (bortezomib, zoledronate) will also be added to culture media in this assay and results compared to previous in vivo studies.

As with murine skeletal samples from tumour-bone disease models, bone from this assay will be assessed by microCT scanning and histomorphometric analysis to quantify bone volume and architecture, bone mineral density, tumour and bone cell distribution. In addition, media will be collected to assess the release of bone- (alkaline- and acid phosphatase) and tumour-derived factors (PTHrP, TGF-B, myeloma-specific IgG2bk) and correlated with measurements obtained using stored serum from mice from past experiments. Representative STANDARD slices from throughout the bone core will generate baseline data with SAMPLE slices cultured under experimental conditions.


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