title: Organotypic slice cultures of human glioblastoma reveal different susceptibilities to treatments.
abstract: BACKGROUND
  Glioblastoma multiforme is the most common lethal brain tumor in human adults, with no major therapeutic breakthroughs in recent decades. Research is based mostly on human tumor cell lines deprived of their organotypic environment or inserted into immune-deficient animals required for graft survival. Here, we describe how glioblastoma specimens obtained from surgical biopsy material can be sectioned and transferred into cultures within minutes.
  
  
  METHODS
  Slices were kept in 6-well plates, allowing direct observation, application of temozolomide, and irradiation. At the end of experiments, slice cultures were processed for histological analysis including hematoxylin-eosin staining, detection of proliferation (Ki67), apoptosis/cell death (cleaved caspase 3, propidium iodide), DNA double-strand breaks (γH2AX), and neural subpopulations. First clinical trials employed irradiation with the heavy ion carbon for the treatment of glioblastoma patients, but the biological effects and most effective dose regimens remain to be established. Therefore, we developed an approach to expose glioblastoma slice cultures to (12)C and X-rays.
  
  
  RESULTS
  We found preservation of the individual histopathology over at least 16 days. Treatments resulted in activation of caspase 3, inhibition of proliferation, and cell loss. Irradiation induced γH2AX. In line with clinical observations, individual tumors differed significantly in their susceptibility to temozolomide (0.4%-2.5% apoptosis and 1%-15% cell loss).
  
  
  CONCLUSIONS
  Glioblastoma multiforme slice cultures provide a unique tool to explore susceptibility of individual tumors for specific therapies including heavy ions, thus potentially allowing more personalized treatments plus exploration of mechanisms of (and strategies to overcome) tumor resistance.
authors.affiliation: Institute of Anatomy, University of Leipzig, Liebigstrasse 13, 04103 Leipzig, Germany. felicitas.merz@medizin.uni-leipzig.de
authors.forename: Felicitas
authors.initials: F
authors.lastname: Merz
authors.forename: Frank
authors.initials: F
authors.lastname: Gaunitz
authors.forename: Faramarz
authors.initials: F
authors.lastname: Dehghani
authors.forename: Christof
authors.initials: C
authors.lastname: Renner
authors.forename: Jürgen
authors.initials: J
authors.lastname: Meixensberger
authors.forename: Angelika
authors.initials: A
authors.lastname: Gutenberg
authors.forename: Alf
authors.initials: A
authors.lastname: Giese
authors.forename: Kosta
authors.initials: K
authors.lastname: Schopow
authors.forename: Christian
authors.initials: C
authors.lastname: Hellwig
authors.forename: Michael
authors.initials: M
authors.lastname: Schäfer
authors.forename: Manfred
authors.initials: M
authors.lastname: Bauer
authors.forename: Horst
authors.initials: H
authors.lastname: Stöcker
authors.forename: Gisela
authors.initials: G
authors.lastname: Taucher-Scholz
authors.forename: Marco
authors.initials: M
authors.lastname: Durante
authors.forename: Ingo
authors.initials: I
authors.lastname: Bechmann
mesh_terms: D000328:Adult; D018906:Antineoplastic Agents, Alkylating / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects / Q000528:radiation effects; D001932:Brain Neoplasms / Q000188:drug therapy / Q000473:pathology* / Q000532:radiotherapy; D002244:Carbon / Q000494:pharmacology; D049109:Cell Proliferation / Q000187:drug effects / Q000528:radiation effects; D053903:DNA Breaks, Double-Stranded / Q000187:drug effects / Q000528:radiation effects; D019175:DNA Methylation / Q000187:drug effects / Q000528:radiation effects; D015254:DNA Modification Methylases / Q000235:genetics; D045643:DNA Repair Enzymes / Q000235:genetics; D003606:Dacarbazine / Q000031:analogs & derivatives* / Q000494:pharmacology; D005909:Glioblastoma / Q000188:drug therapy / Q000473:pathology* / Q000532:radiotherapy; D006801:Humans; D009924:Organ Culture Techniques; D011401:Promoter Regions, Genetic / Q000235:genetics; D000077204:Temozolomide; D014407:Tumor Cells, Cultured; D025521:Tumor Suppressor Proteins / Q000235:genetics; D014965:X-Rays
chemical_list: D018906:Antineoplastic Agents, Alkylating; D025521:Tumor Suppressor Proteins; D002244:Carbon; D003606:Dacarbazine; D015254:DNA Modification Methylases; C515491:MGMT protein, human; D045643:DNA Repair Enzymes; D000077204:Temozolomide
keywords: glioblastoma multiforme; heavy ions; human test system; organotypic slice culture