Title |
Short term culture of breast cancer tissues to study the activity of the anticancer drug taxol in an intact tumor environment
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Published in |
BMC Cancer, April 2006
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DOI | 10.1186/1471-2407-6-86 |
Pubmed ID | |
Authors |
Heiko van der Kuip, Thomas E Mürdter, Maike Sonnenberg, Monika McClellan, Susanne Gutzeit, Andreas Gerteis, Wolfgang Simon, Peter Fritz, Walter E Aulitzky |
Abstract |
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. 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. 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. 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. |
Mendeley readers
Geographical breakdown
Country | Count | As % |
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France | 2 | 1% |
United States | 2 | 1% |
Italy | 1 | <1% |
United Kingdom | 1 | <1% |
South Africa | 1 | <1% |
Unknown | 131 | 95% |
Demographic breakdown
Readers by professional status | Count | As % |
---|---|---|
Researcher | 40 | 29% |
Student > Ph. D. Student | 30 | 22% |
Student > Master | 15 | 11% |
Professor > Associate Professor | 10 | 7% |
Student > Doctoral Student | 9 | 7% |
Other | 20 | 14% |
Unknown | 14 | 10% |
Readers by discipline | Count | As % |
---|---|---|
Agricultural and Biological Sciences | 40 | 29% |
Biochemistry, Genetics and Molecular Biology | 25 | 18% |
Medicine and Dentistry | 23 | 17% |
Engineering | 11 | 8% |
Chemistry | 5 | 4% |
Other | 10 | 7% |
Unknown | 24 | 17% |