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

• Published in: Scientific reports 2015-12, Vol.5 (1), p.17187-17187, Article 17187
• Main Authors: Davies, Emma J., Dong, Meng, Gutekunst, Matthias, Närhi, Katja, van Zoggel, Hanneke J. A. A., Blom, Sami, Nagaraj, Ashwini, Metsalu, Tauno, Oswald, Eva, Erkens-Schulze, Sigrun, Delgado San Martin, Juan A., Turkki, Riku, Wedge, Stephen R., af Hällström, Taija M., Schueler, Julia, van Weerden, Wytske M., Verschuren, Emmy W., Barry, Simon T., van der Kuip, Heiko, Hickman, John A.
• Format: Article
• Language: English
• Subjects: 13; 13/105; 13/106; 14; 38; 38/77; 631/67/2329; 631/67/70; Animal models; Animals; Biomarkers; Cell Line, Tumor; Cellular stress response; Cultivation; DNA microarrays; Gene Expression; Heterografts; Humanities and Social Sciences; Humans; Immunohistochemistry - methods; Mice; multidisciplinary; Neoplasms - genetics; Neoplasms - metabolism; Neoplasms - pathology; Oxygen - metabolism; Principal Component Analysis; Protein expression; Questioning; Real-Time Polymerase Chain Reaction; Science; Signal Transduction; Stress, Physiological; Tissue Array Analysis; Tissue Culture Techniques; Transcription; Tumors
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep17187

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.