Organotypic slice cultures of human gastric and esophagogastric junction cancer

Abstract

Gastric and esophagogastric junction cancers are heterogeneous and aggressive tumors with an unpredictable response to cytotoxic treatment. New methods allowing for the analysis of drug resistance are needed. Here, we describe a novel technique by which human tumor specimens can be cultured ex vivo, preserving parts of the natural cancer microenvironment. Using a tissue chopper, fresh surgical tissue samples were cut in 400 μm slices and cultivated in 6‐well plates for up to 6 days. The slices were processed for routine histopathology and immunohistochemistry. Cytokeratin stains (CK8, AE1/3) were applied for determining tumor cellularity, Ki‐67 for proliferation, and cleaved caspase‐3 staining for apoptosis. The slices were analyzed under naive conditions and following 2–4 days in vitro exposure to 5‐FU and cisplatin. The slice culture technology allowed for a good preservation of tissue morphology and tumor cell integrity during the culture period. After chemotherapy exposure, a loss of tumor cellularity and an increase in apoptosis were observed. Drug sensitivity of the tumors could be assessed. Organotypic slice cultures of gastric and esophagogastric junction cancers were successfully established. Cytotoxic drug effects could be monitored. They may be used to examine mechanisms of drug resistance in human tissue and may provide a unique and powerful ex vivo platform for the prediction of treatment response.

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

“…NSCLC slices cultured on rotating incubation units, to ensure continuous oxygenation, were fragile and showed dynamic culture‐induced changes, most prominently altered proliferation and mTORC1 hyperactivation. Although previous reports showed sustained viability of clinical tumour slices for 4–14 days , these studies derived conclusions from the selective analysis of cultured slices, whereas we systematically compared neighbouring cultured and uncultured slices. Furthermore, our study is the first to deeply analyse intratumour oncogenic signalling, and shows that signalling and proliferation alterations take place prior to overt changes in viability.…”

“…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 . However, the question remains of how the in situ spatial signalling heterogeneity affects the pharmacodynamics of signalling inhibitors.…”

Helicobacter pylori Infection Modulates Host Cell Metabolism through VacA-Dependent Inhibition of mTORC1

“…2B). Finally, we observed VacA-dependent inhibition of mTORC1 signaling activity within an ex vivo gastric tissue slice model derived from the stomachs of C57BL/6J mice (Koerfer et al, 2016) (Fig. 2C).…”

Ex vivo treatment of prostate tumor tissue recapitulates in vivo therapy response

“…As an alternative, ex vivo culturing of tumor slices represent a solid model system for drug sensitivity testing due to its relatively short generation time and reflection of the tumor microenvironment . For example, several studies have shown that breast tumor tissue slices can be used to assess chemotherapy response in the context of personalized medicine .…”