title: Capturing complex tumour biology in vitro: histological and molecular characterisation of precision cut slices.
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.
authors.affiliation: Oncology iMed, Bioscience, AstraZeneca, Alderley Park, Macclesfield, SK10 4TG, United Kingdom.
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Davies
authors.affiliation: Dr Margarete Fischer-Bosch Institute of Clinical Pharmacology and University of Tuebingen, Auerbachstrasse 112, 70376 Stuttgart, Germany.
authors.forename: Meng
authors.initials: M
authors.lastname: Dong
authors.affiliation: Dr Margarete Fischer-Bosch Institute of Clinical Pharmacology and University of Tuebingen, Auerbachstrasse 112, 70376 Stuttgart, Germany.
authors.forename: Matthias
authors.initials: M
authors.lastname: Gutekunst
authors.affiliation: FIMM, University of Helsinki; Helsinki, Finland.
authors.forename: Katja
authors.initials: K
authors.lastname: Närhi
authors.affiliation: Department of Urology, Erasmus MC Rotterdam, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Hanneke J A A
authors.initials: HJ
authors.lastname: van Zoggel
authors.affiliation: FIMM, University of Helsinki; Helsinki, Finland.
authors.forename: Sami
authors.initials: S
authors.lastname: Blom
authors.affiliation: FIMM, University of Helsinki; Helsinki, Finland.
authors.forename: Ashwini
authors.initials: A
authors.lastname: Nagaraj
authors.affiliation: Institute of Computer Science, University of Tartu, J. Liivi 2, 50409, Tartu, Estonia.
authors.forename: Tauno
authors.initials: T
authors.lastname: Metsalu
authors.affiliation: Oncotest GmbH, Am Flughafen 12-14, 79108 Freiburg, Germany.
authors.forename: Eva
authors.initials: E
authors.lastname: Oswald
authors.affiliation: Department of Urology, Erasmus MC Rotterdam, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Sigrun
authors.initials: S
authors.lastname: Erkens-Schulze
authors.affiliation: Oncology iMed, Bioscience, AstraZeneca, Alderley Park, Macclesfield, SK10 4TG, United Kingdom.
authors.forename: Juan A
authors.initials: JA
authors.lastname: Delgado San Martin
authors.affiliation: FIMM, University of Helsinki; Helsinki, Finland.
authors.forename: Riku
authors.initials: R
authors.lastname: Turkki
authors.affiliation: Oncology iMed, Bioscience, AstraZeneca, Alderley Park, Macclesfield, SK10 4TG, United Kingdom.
authors.forename: Stephen R
authors.initials: SR
authors.lastname: Wedge
authors.affiliation: FIMM, University of Helsinki; Helsinki, Finland.
authors.forename: Taija M
authors.initials: TM
authors.lastname: af Hällström
authors.affiliation: Oncotest GmbH, Am Flughafen 12-14, 79108 Freiburg, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Schueler
authors.affiliation: Department of Urology, Erasmus MC Rotterdam, PO Box 2040, 3000 CA Rotterdam, The Netherlands.
authors.forename: Wytske M
authors.initials: WM
authors.lastname: van Weerden
authors.affiliation: FIMM, University of Helsinki; Helsinki, Finland.
authors.forename: Emmy W
authors.initials: EW
authors.lastname: Verschuren
authors.affiliation: Oncology iMed, Bioscience, AstraZeneca, Alderley Park, Macclesfield, SK10 4TG, United Kingdom.
authors.forename: Simon T
authors.initials: ST
authors.lastname: Barry
authors.affiliation: Dr Margarete Fischer-Bosch Institute of Clinical Pharmacology and University of Tuebingen, Auerbachstrasse 112, 70376 Stuttgart, Germany.
authors.forename: Heiko
authors.initials: H
authors.lastname: van der Kuip
authors.affiliation: Institut de Recherches Servier c/o 126 boulevard Pereire, 75017 Paris, France.
authors.forename: John A
authors.initials: JA
authors.lastname: Hickman
mesh_terms: D000818:Animals; D015415:Biomarkers; D045744:Cell Line, Tumor; D015870:Gene Expression; D064593:Heterografts; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D009369:Neoplasms / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology*; D010100:Oxygen / Q000378:metabolism; D025341:Principal Component Analysis; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D013312:Stress, Physiological; D046888:Tissue Array Analysis; D046509:Tissue Culture Techniques
chemical_list: D015415:Biomarkers; D010100:Oxygen