Establishment and characterization of primary glioblastoma cell lines from fresh and frozen material: a detailed comparison

PLoS One. 2013 Aug 7;8(8):e71070. doi: 10.1371/journal.pone.0071070. eCollection 2013.

Abstract

Background: Development of clinically relevant tumor model systems for glioblastoma multiforme (GBM) is important for advancement of basic and translational biology. High molecular heterogeneity of GBM tumors is well recognized, forming the rationale for molecular tests required before administration of several of the novel therapeutics rapidly entering the clinics. One model that has gained wide acceptance is the primary cell culture model. The laborious and time consuming process is rewarded with a relative high success rate (about 60%). We here describe and evaluate a very simple cryopreservation procedure for GBM tissue prior to model establishment that will considerably reduce the logistic complexity.

Methods: Twenty-seven GBM samples collected ad hoc were prepared for primary cell culture freshly from surgery (#1) and after cryopreservation (#2).

Results: Take rates after cryopreservation (59%) were as satisfactory as from fresh tissue (63%; p = 1.000). We did not observe any relevant molecular or phenotypic differences between cell lines established from fresh or vitally frozen tissue. Further, sensitivity both towards standard chemotherapeutic agents (Temozolomide, BCNU and Vincristine) and novel agents like the receptor tyrosine kinase inhibitor Imatinib did not differ.

Conclusions: Our simple cryopreservation procedure facilitates collection, long-time storage and propagation (modeling) of clinical GBM specimens (potentially also from distant centers) for basic research, (pre-) clinical studies of novel therapies and individual response prediction.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Aged
  • Aged, 80 and over
  • Antineoplastic Agents / pharmacology
  • Benzamides / pharmacology
  • Brain Neoplasms / genetics
  • Brain Neoplasms / metabolism
  • Brain Neoplasms / pathology*
  • Carmustine / pharmacology
  • Cell Line, Tumor
  • Cell Proliferation*
  • Cell Survival / drug effects
  • Cryopreservation / methods*
  • Dacarbazine / analogs & derivatives
  • Dacarbazine / pharmacology
  • Dose-Response Relationship, Drug
  • ErbB Receptors / genetics
  • Female
  • Gene Dosage
  • Glial Fibrillary Acidic Protein / metabolism
  • Glioblastoma / genetics
  • Glioblastoma / metabolism
  • Glioblastoma / pathology*
  • Humans
  • Imatinib Mesylate
  • Male
  • Middle Aged
  • Piperazines / pharmacology
  • Protein Kinase Inhibitors / pharmacology
  • Pyrimidines / pharmacology
  • Temozolomide
  • Tumor Cells, Cultured
  • Vincristine / pharmacology

Substances

  • Antineoplastic Agents
  • Benzamides
  • Glial Fibrillary Acidic Protein
  • Piperazines
  • Protein Kinase Inhibitors
  • Pyrimidines
  • Vincristine
  • Dacarbazine
  • Imatinib Mesylate
  • EGFR protein, human
  • ErbB Receptors
  • Carmustine
  • Temozolomide

Grants and funding

This work was supported by the “Wilhelm Vaillant Stiftung”, the “Verein zur Förderung krebskranker Kinder Rostock, e.V.” and a founding program of the university medicine Rostock (FORUN). The state Mecklenburg-Vorpommern granted CSM a graduate scholarship to perform this work. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.