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In Vitro Cell Dev Biol Anim. 2009 Sep;45(8):442-50. doi: 10.1007/s11626-009-9212-7. Epub 2009 Jun 16.

Precision-cut slice cultures of tumors from MMTV-neu mice for the study of the ex vivo response to cytokines and cytotoxic drugs.

Author information

1
Division of Medical Biochemistry, Biocenter, Innsbruck Medical University, Innsbruck, Austria.

Abstract

Ex vivo analysis of signaling pathways operating in tumor tissue is complicated by the three-dimensional structure, in particular by stroma-epithelial interactions. Studies performed with pure populations of tumor cells usually do not take into account this issue. One possibility to preserve the tissue architecture is the use of tumor slices. However, diffusion of oxygen and nutrients may become limiting factors, resulting in decreased cell viability and change of tissue morphology, especially after long-term incubation of slices. By using precision cut slices of defined thickness, we were able to establish culture conditions for tumor material obtained from MMTV-neu transgenic mice, which allow the study of the action of cytokines and cytotoxic drugs for up to 24 h. A slice thickness of 160 mum was found to be optimal for viability and handling of material. These slices were highly responsive to the action of the cytokine IFN-gamma, as evident form the increase of pY701 STAT1, detected by both immunohistochemistry and western blotting, and by the increase of mRNA levels of the IFN-gamma response genes IRF-1, SOCS-1, and STAT1, analyzed by reverse transcriptase-polymerase chain reaction. Furthermore, induction of apoptosis and increase of DNA damage could be monitored after treatment with IFN-gamma or doxorubicin. The slices were also a convenient source for the establishment of explant cultures of tumor epithelial cells. It is concluded that cultivation of precision-cut tumor slices provides a convenient way for the ex vivo molecular analysis of MMTV-neu tumor tissue under conditions which closely simulate the situation in vivo and can provide an alternative to in vivo experiments.

PMID:
19533258
DOI:
10.1007/s11626-009-9212-7
[Indexed for MEDLINE]

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