Book of abstract 2008

Development of three-dimensional (3D) all-human, in vitro models for
study of the biology of primary and metastatic brain tumours
S. A. Murray, K. Fry, G. J. Pilkington
Cellular & Molecular Neuro-oncology Group, Institute of Biomedical & Biomolecular Sciences, School
of Pharmacy & Biomedical Sciences, University of Portsmouth, St Michael’s Building, White Swan Road,
Portsmouth PO1 2DT UK
Animal models for the study of local invasive behaviour and metastasis of brain tumour
poorly reflect the situation in human counterpart neoplasms. Moreover, 3D in vitro
invasion models for invasion generally utilise non-human (rat/mouse) glioma cells and/
or rat brain/chick heart fragments as a “target” for invasion. Similarly, in vitro models
of the B-BB generally utilise porcine or murine brain endothelium and rat astrocytes. In
addition, these models are grown in foetal calf serum supplemented conditions which
modify growth rates and cell adhesive properties. Our aims were, therefore, to develop 3D
in vitro models from human brain-derived cells for the study of: a) brain cell:glioma cell
interaction and invasive behaviour and b) passage of metastatic somatic cancer cells across
the cellular blood-brain barrier (B-BB).
For the invasion model we have used human glioma biopsy-derived cells and temporal
lobe epilepsy resection (TLER) brain maintained as spheroids through an adaptation
of the hanging drop method and juxtaposed in 3D confrontation culture under human
serum (HS) supplementation. For the B-BB model we have used astrocyte-rich human
cultures from TELR brain in combination with human cerebral microvascular endothelial
cells (CMECs) immortalised with hTERT/SV40LargeT, under HS supplementation. All
cells have been characterised with appropriate immuno markers using flow cytometry
and immunocytochemistry while growth curves and adhesion properties have been
established.
Growth curves, spheroid growth rates, scanning electron microscopy, confocal microscopy,
antigenic expression, adhesive properties and growth on Transwells have been established
for each model and for the individual component cells.
We are currently
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assessing the models using a combination of live cell imaging, WetSEM,
TIRF microscopy and confocal microscopy in studies where putative inhibitors of
invasion/metastasis are under investigation.
This research is supported by grant funding from the Dr Hadwen Trust (www.drhadwentrust.co.uk) and
the Lord Dowding Fund and a PhD bursary from the Institute of Biomedical & Biomolecular Sciences,
University of Portsmouth.
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