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Tumor Biology

, Volume 35, Issue 7, pp 6221–6233 | Cite as

Characterization of the 9L gliosarcoma implanted in the Fischer rat: an orthotopic model for a grade IV brain tumor

  • Audrey Bouchet
  • Marie Bidart
  • Imen Miladi
  • Céline Le Clec’h
  • Raphaël Serduc
  • Charles Coutton
  • Pierrick Regnard
  • Enam Khalil
  • Sandrine Dufort
  • Benjamin Lemasson
  • Jean Laissue
  • Laurent Pelletier
  • Géraldine Le DucEmail author
Review

Abstract

Among rodent models for brain tumors, the 9L gliosarcoma is one of the most widely used. Our 9L-European Synchrotron Radiation Facility (ESRF) model was developed from cells acquired at the Brookhaven National Laboratory (NY, USA) in 1997 and implanted in the right caudate nucleus of syngeneic Fisher rats. It has been largely used by the user community of the ESRF during the last decade, for imaging, radiotherapy, and chemotherapy, including innovative treatments based on particular irradiation techniques and/or use of new drugs. This work presents a detailed study of its characteristics, assessed by magnetic resonance imaging (MRI), histology, immunohistochemistry, and cytogenetic analysis. The data used for this work were from rats sampled in six experiments carried out over a 3-year period in our lab (total number of rats = 142). The 9L-ESRF tumors were induced by a stereotactic inoculation of 104 9L cells in the right caudate nucleus of the brain. The assessment of vascular parameters was performed by MRI (blood volume fraction and vascular size index) and by immunostaining of vessels (rat endothelial cell antigen-1 and type IV collagen). Immunohistochemistry and regular histology were used to describe features such as tumor cell infiltration, necrosis area, nuclear pleomorphism, cellularity, mitotic characteristics, leukocytic infiltration, proliferation, and inflammation. Moreover, for each of the six experiments, the survival of the animals was assessed and related to the tumor growth observed by MRI or histology. Additionally, the cytogenetic status of the 9L cells used at ESRF lab was investigated by comparative genomics hybridization analysis. Finally, the response of the 9L-ESRF tumor to radiotherapy was estimated by plotting the survival curves after irradiation. The median survival time of 9L-ESRF tumor-bearing rats was highly reproducible (19–20 days). The 9L-ESRF tumors presented a quasi-exponential growth, were highly vascularized with a high cellular density and a high proliferative index, accompanied by signs of inflammatory responses. We also report an infiltrative pattern which is poorly observed on conventional 9 L tumor. The 9L-ESRF cells presented some cytogenetic specificities such as altered regions including CDK4, CDKN2A, CDKN2B, and MDM2 genes. Finally, the lifespan of 9L-ESRF tumor-bearing rats was enhanced up to 28, 35, and 45 days for single doses of 10, 20, and 2 × 20 Gy, respectively. First, this report describes an animal model that is used worldwide. Second, we describe few features typical of our model if compared to other 9L models worldwide. Altogether, the 9L-ESRF tumor model presents characteristics close to the human high-grade gliomas such as high proliferative capability, high vascularization and a high infiltrative pattern. Its response to radiotherapy demonstrates its potential as a tool for innovative radiotherapy protocols.

Keywords

Glioma 9L Rat Infiltration Hypervascularization Radiotherapy Cytogenetic 

Abbreviations

BVf

Blood volume fraction

CGH

Comparative genomics hybridization

DNA

Deoxyribonucleic acid

ECACC

European Collection of Cell Cultures

EGFR

Epidermal growth factor receptor

ESRF

European Synchrotron Radiation Facility

GFAP

Glial fibrillary acidic protein

MRI

Magnetic resonance imaging

MST

Median survival time

PDGFR

Platelet-derived growth factor receptor

SEM

Standard error to the mean

VSI

Vessel size index

Notes

Acknowledgments

We thank Charlène Caloud, Hélène Bernard, and Dominique Dalléry for their technical help.

Conflicts of interest

None

Financial support

This project was partially granted by the Institut National du Cancer of France (Bioresys project #200187).

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  • Audrey Bouchet
    • 1
    • 2
    • 7
  • Marie Bidart
    • 1
    • 8
  • Imen Miladi
    • 3
    • 9
  • Céline Le Clec’h
    • 2
    • 10
  • Raphaël Serduc
    • 2
    • 11
  • Charles Coutton
    • 4
  • Pierrick Regnard
    • 2
    • 12
  • Enam Khalil
    • 2
    • 13
  • Sandrine Dufort
    • 5
    • 6
  • Benjamin Lemasson
    • 1
  • Jean Laissue
    • 7
  • Laurent Pelletier
    • 1
    • 8
  • Géraldine Le Duc
    • 2
    • 14
    Email author
  1. 1.Grenoble Institut des NeurosciencesUniversité Joseph Fourier, INSERM U836La Tronche CedexFrance
  2. 2.Biomedical BeamlineEuropean Synchrotron Radiation FacilityGrenoble CedexFrance
  3. 3.Laboratoire de Physico-Chimie des Matériaux Luminescents, UMR 5620 CNRS – UCBLUniversité de LyonVilleurbanne CedexFrance
  4. 4.Département Génétique et Procréation, Laboratoire de Génétique Chromosomique, Hôpital Couple-EnfantCHU Grenoble, Equipe GIT, UJF/CNRS AGIM FRE 34056La TroncheFrance
  5. 5.Nano-H SASSaint-Quentin FallavierFrance
  6. 6.INSERM, CRI, U823, Institut Albert BonniotGrenobleFrance
  7. 7.Institute of AnatomyUniversity of BernBernSwitzerland
  8. 8.Grenoble University HospitalGrenoble CedexFrance
  9. 9.INSERM, Imagerie Moléculaire et thérapie Vectorisée, UMR 990, Université d’AuvergneClermont-FerrandFrance
  10. 10.CEA, LETI, CLINATECGrenobleFrance
  11. 11.INSERM, U836GrenobleFrance
  12. 12.Plateforme SILABE-ADUEIS, Fort FochNiederhausbergenFrance
  13. 13.Faculty of Pharmacy, University of JordanAmmanJordan
  14. 14.ESRFGrenobleFrance

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