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See 1 citation in BMC Cancer 2006:

BMC Cancer. 2006 Apr 7;6:86.

Short term culture of breast cancer tissues to study the activity of the anticancer drug taxol in an intact tumor environment.

Author information

1
Dr, Margarete Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart, Germany. heiko.van-der-kuip@ikp-stuttgart.de

Abstract

BACKGROUND:

Sensitivity of breast tumors to anticancer drugs depends upon dynamic interactions between epithelial tumor cells and their microenvironment including stromal cells and extracellular matrix. To study drug-sensitivity within different compartments of an individual tumor ex vivo, culture models directly established from fresh tumor tissues are absolutely essential.

METHODS:

We prepared 0.2 mm thick tissue slices from freshly excised tumor samples and cultivated them individually in the presence or absence of taxol for 4 days. To visualize viability, cell death, and expression of surface molecules in different compartments of non-fixed primary breast cancer tissues we established a method based on confocal imaging using mitochondria- and DNA-selective dyes and fluorescent-conjugated antibodies. Proliferation and apoptosis was assessed by immunohistochemistry in sections from paraffin-embedded slices. Overall viability was also analyzed in homogenized tissue slices by a combined ATP/DNA quantification assay.

RESULTS:

We obtained a mean of 49 tissue slices from 22 breast cancer specimens allowing a wide range of experiments in each individual tumor. In our culture system, cells remained viable and proliferated for at least 4 days within their tissue environment. Viability of tissue slices decreased significantly in the presence of taxol in a dose-dependent manner. A three-color fluorescence viability assay enabled a rapid and authentic estimation of cell viability in the different tumor compartments within non-fixed tissue slices.

CONCLUSION:

We describe a tissue culture method combined with a novel read out system for both tissue cultivation and rapid assessment of drug efficacy together with the simultaneous identification of different cell types within non-fixed breast cancer tissues. This method has potential significance for studying tumor responses to anticancer drugs in the complex environment of a primary cancer tissue.

PMID:
16603054
PMCID:
PMC1456977
DOI:
10.1186/1471-2407-6-86
[Indexed for MEDLINE]
Free PMC Article

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