Capturing complex tumour biology in vitro: histological and molecular characterisation of precision cut slices.

Emma J Davies, Meng Dong, Matthias Gutekunst, Katja Närhi, Hanneke J A A van Zoggel, Sami Blom, Ashwini Nagaraj, Tauno Metsalu, Eva Oswald, Sigrun Erkens-Schulze, Juan A Delgado San Martin, Riku Turkki, Stephen R Wedge, Taija M af Hällström, Julia Schueler, Wytske M van Weerden, Emmy W Verschuren, Simon T Barry, Heiko van der Kuip, John A Hickman

Index: Sci. Rep. 5 , 17187, (2015)

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Abstract

Precision-cut slices of in vivo tumours permit interrogation in vitro of heterogeneous cells from solid tumours together with their native microenvironment. They offer a low throughput but high content in vitro experimental platform. Using mouse models as surrogates for three common human solid tumours, we describe a standardised workflow for systematic comparison of tumour slice cultivation methods and a tissue microarray-based method to archive them. Cultivated slices were compared to their in vivo source tissue using immunohistochemical and transcriptional biomarkers, particularly of cellular stress. Mechanical slicing induced minimal stress. Cultivation of tumour slices required organotypic support materials and atmospheric oxygen for maintenance of integrity and was associated with significant temporal and loco-regional changes in protein expression, for example HIF-1α. We recommend adherence to the robust workflow described, with recognition of temporal-spatial changes in protein expression before interrogation of tumour slices by pharmacological or other means.