title: Short term culture of breast cancer tissues to study the activity of the anticancer drug taxol in an intact tumor environment.
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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Dr, Margarete Fischer-Bosch Institute of Clinical Pharmacology, Stuttgart, Germany. heiko.van-der-kuip@ikp-stuttgart.de
authors.forename: Heiko
authors.initials: H
authors.lastname: van der Kuip
authors.forename: Thomas E
authors.initials: TE
authors.lastname: Mürdter
authors.forename: Maike
authors.initials: M
authors.lastname: Sonnenberg
authors.forename: Monika
authors.initials: M
authors.lastname: McClellan
authors.forename: Susanne
authors.initials: S
authors.lastname: Gutzeit
authors.forename: Andreas
authors.initials: A
authors.lastname: Gerteis
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Simon
authors.forename: Peter
authors.initials: P
authors.lastname: Fritz
authors.forename: Walter E
authors.initials: WE
authors.lastname: Aulitzky
mesh_terms: D002288:Adenocarcinoma, Mucinous / Q000473:pathology*; D000255:Adenosine Triphosphate / Q000032:analysis; D000972:Antineoplastic Agents, Phytogenic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001943:Breast Neoplasms / Q000473:pathology*; D018270:Carcinoma, Ductal, Breast / Q000473:pathology*; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival; D004273:DNA, Neoplasm / Q000032:analysis; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005260:Female; D006801:Humans; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence; D017239:Paclitaxel / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000737:chemistry / Q000187:drug effects
chemical_list: D000972:Antineoplastic Agents, Phytogenic; D004273:DNA, Neoplasm; D000255:Adenosine Triphosphate; D017239:Paclitaxel