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.

Spatial aspects of oncogenic signalling determine the response to combination therapy in slice explants from Kras‐driven lung tumours

“…Murine NSCLC tumours and clinical specimens were precision‐cut to make 200‐μm slices with a VT1200S microtome (Leica Biosystems, Wetzlar, Germany), as described previously . Murine slices were transferred to rotating incubators (Alabama Research & Development, Munford, AL, USA) (supplementary material, Figure S2A), within 90–120 min after mice had been killed.…”

“…Precision‐cut tumour slices, which capture the native tumour in its microenvironment, constitute an attractive model with which to address this need. Within the IMI‐PREDECT project (http://www.predect.eu), we developed a workflow for short‐term culture of tumour slices, and showed that organotypic supports and atmospheric oxygen are strict requirements for tissue viability . A number of other studies have shown that responses to cytotoxic agents or selected targeted compounds can be modelled in clinical tumour‐derived slices .…”

The Synthetic β-Nitrostyrene Derivative CYT-Rx20 Inhibits Esophageal Tumor Growth and Metastasis via PI3K/AKT and STAT3 Pathways

“…The results for Ki-67, phospho-AKT, and phospho-STAT3 staining were determined by two independent experts under the same condition. For quantification, the sections of each immunostaining were scored using the method of histochemical score (H-score), which was calculated as the product of percentage of stained cells and intensity of staining [22,23]. …”

Tumour-on-a-chip: microfluidic models of tumour morphology, growth and microenvironment

“…Primary tumour tissues from biopsy or surgical resection can be embedded in ECM and cultured as an in vitro model [30][31][32][33][34] (figure 2a). The embedded tumour sections retain the tumour vasculature, nearby stroma and the heterogeneity of the tumour cells.…”