Journal of Neuro-Oncology

, Volume 106, Issue 3, pp 493–504 | Cite as

Isolation of a new cell population in the glioblastoma microenvironment

  • Anne Clavreul
  • Amandine Etcheverry
  • Agnès Chassevent
  • Véronique Quillien
  • Tony Avril
  • Marie-Lise Jourdan
  • Sophie Michalak
  • Patrick François
  • Jean-Luc Carré
  • Jean Mosser
  • The Grand Ouest Glioma Project Network
  • Philippe Menei
Laboratory Investigation - Human/Animal Tissue


Glioblastoma (GB) is a highly infiltrative tumor recurring in 90% of cases within a few centimeters of the resection cavity, even in cases of complete tumor resection and adjuvant chemo/radiotherapy. This observation highlights the importance of understanding this special zone of brain tissue surrounding the tumor. It is becoming clear that the nonneoplastic stromal compartment of most solid cancers plays an active role in tumor proliferation, invasion, and metastasis. Very little information, other than that concerning angiogenesis and immune cells, has been collected for stromal cells from GB. As part of a translational research program, we have isolated a new stromal cell population surrounding GB by computer-guided stereotaxic biopsies and primary culture. We named these cells GB-associated stromal cells (GASCs). GASCs are diploid, do not display the genomic alterations typical of GB cells, and have phenotypic and functional properties in common with the cancer-associated fibroblasts (CAFs) described in the stroma of carcinomas. In particular, GASCs express markers associated with CAFs such as fibroblast surface protein, alpha-smooth muscle actin (α-SMA), and platelet-derived growth factor receptor-beta (PDGFRβ). Furthermore, GASCs have a molecular expression profile different from that of control stromal cells derived from non-GB peripheral brain tissues. GASCs were also found to have tumor-promoting effects on glioma cells in vitro and in vivo. The isolation of GASCs in a tumor of neuroepithelial origin was unexpected, and further studies are required to determine their potential as a target for antiglioma treatment.


Translational study Glioblastoma Microenvironment Cancer-associated fibroblasts Primary cultures 



Cancer-associated fibroblasts


Dulbecco’s modified Eagle’s medium


Absolute fold change


GB-associated stromal cells




Human AB serum


Human mesenchymal stem cells


Interface zone


Necrotic zone


Peripheral brain zone


Platelet-derived growth factor receptor-beta


Real-time quantitative polymerase chain reaction


Alpha-smooth muscle actin


Tumor zone


  1. 1.
    Stupp R, Hegi ME, Mason WP et al (2009) Effects of radiotherapy with concomitant and adjuvant temozolomide versus radiotherapy alone on survival in glioblastoma in a randomised phase III study: 5-year analysis of the EORTC–NCIC trial. Lancet Oncol 10:459–466PubMedCrossRefGoogle Scholar
  2. 2.
    Giese A, Kucinski T, Knopp U et al (2004) Pattern of recurrence following local chemotherapy with biodegradable carmustine (BCNU) implants in patients with glioblastoma. J Neurooncol 66:351–360PubMedCrossRefGoogle Scholar
  3. 3.
    Lefranc F, Brotchi J, Kiss R (2005) Possible future issues in the treatment of glioblastomas: special emphasis on cell migration and the resistance of migrating glioblastoma cells to apoptosis. J Clin Oncol 23:2411–2422PubMedCrossRefGoogle Scholar
  4. 4.
    Liang BC, Thornton AF Jr, Sandler HM et al (1991) Malignant astrocytomas: focal tumor recurrence after focal external beam radiation therapy. J Neurosurg 75:559–563PubMedCrossRefGoogle Scholar
  5. 5.
    Tlsty TD, Coussens LM (2006) Tumor stroma and regulation of cancer development. Annu Rev Pathol 1:119–150PubMedCrossRefGoogle Scholar
  6. 6.
    Ostman A, Augsten M (2009) Cancer-associated fibroblasts and tumor growth-bystanders turning into key players. Curr Opin Genet Dev 19:67–73PubMedCrossRefGoogle Scholar
  7. 7.
    Bhowmick NA, Neilson EG, Moses HL (2004) Stromal fibroblasts in cancer initiation and progression. Nature 432:332–337PubMedCrossRefGoogle Scholar
  8. 8.
    Shimoda M, Mellody KT, Orimo A (2010) Carcinoma-associated fibroblasts are a rate-limiting determinant for tumour progression. Semin Cell Dev Biol 21:19–25PubMedCrossRefGoogle Scholar
  9. 9.
    Franco OE, Shaw AK, Strand DW et al (2010) Cancer associated fibroblasts in cancer pathogenesis. Semin Cell Dev Biol 21:33–39PubMedCrossRefGoogle Scholar
  10. 10.
    Xie Z (2009) Brain tumor stem cells. Neurochem Res 34:2055–2066PubMedCrossRefGoogle Scholar
  11. 11.
    Chen R, Nishimura MC, Bumbaca SM et al (2010) A hierarchy of self-renewing tumor-initiating cell types in glioblastoma. Cancer Cell 17:362–375PubMedCrossRefGoogle Scholar
  12. 12.
    Jain RK, di Tomaso E, Duda DG et al (2007) Angiogenesis in brain tumours. Nat Rev Neurosci 8:610–622PubMedCrossRefGoogle Scholar
  13. 13.
    Yang I, Han SJ, Kaur G et al (2010) The role of microglia in central nervous system immunity and glioma immunology. J Clin Neurosci 17:6–10PubMedCrossRefGoogle Scholar
  14. 14.
    Sonabend AM, Rolle CE, Lesniak MS (2008) The role of regulatory T cells in malignant glioma. Anticancer Res 28:1143–1150PubMedGoogle Scholar
  15. 15.
    Louis DN, Ohgaki H, Wiestler OD et al (2007) The 2007 WHO classification of tumours of the central nervous system. Acta Neuropathol 114:97–109PubMedCrossRefGoogle Scholar
  16. 16.
    Vindelov LL, Christensen IJ, Nissen NI (1983) A detergent-trypsin method for the preparation of nuclei for flow cytometric DNA analysis. Cytometry 3:323–327PubMedCrossRefGoogle Scholar
  17. 17.
    Roger M, Clavreul A, Venier-Julienne MC et al (2010) Mesenchymal stem cells as cellular vehicles for delivery of nanoparticles to brain tumors. Biomaterials 31:8393–8401PubMedCrossRefGoogle Scholar
  18. 18.
    Hupe P, Stransky N, Thiery JP et al (2004) Analysis of array CGH data: from signal ratio to gain and loss of DNA regions. Bioinformatics 20:3413–3422PubMedCrossRefGoogle Scholar
  19. 19.
    Clavreul A, Jean I, Preisser L et al (2009) Human glioma cell culture: two FCS-free media could be recommended for clinical use in immunotherapy. In Vitro Cell Dev Biol Anim 45:500–511PubMedCrossRefGoogle Scholar
  20. 20.
    Glas M, Rath BH, Simon M et al (2010) Residual tumor cells are unique cellular targets in glioblastoma. Ann Neurol 68:264–269PubMedGoogle Scholar
  21. 21.
    Westermark B, Ponten J, Hugosson R (1973) Determinants for the establishment of permanent tissue culture lines from human gliomas. Acta Pathol Microbiol Scand A 81:791–805PubMedGoogle Scholar
  22. 22.
    Rutka JT, Giblin JR, Dougherty DY et al (1987) Establishment and characterization of five cell lines derived from human malignant gliomas. Acta Neuropathol 75:92–103PubMedCrossRefGoogle Scholar
  23. 23.
    Gibbons HM, Hughes SM, Van Roon-Mom W et al (2007) Cellular composition of human glial cultures from adult biopsy brain tissue. J Neurosci Methods 166:89–98PubMedCrossRefGoogle Scholar
  24. 24.
    Orimo A, Weinberg RA (2007) Heterogeneity of stromal fibroblasts in tumors. Cancer Biol Ther 6:618–619PubMedGoogle Scholar
  25. 25.
    Raica M, Cimpean AM (2010) Platelet-derived growth factor (PDGF)/PDGF receptors (PDGFR) axis as target for antitumor and antiangiogenic therapy. Pharmaceuticals 3:572–599CrossRefGoogle Scholar
  26. 26.
    Amillet JM, Ferbus D, Real FX et al (2006) Characterization of human Rab20 overexpressed in exocrine pancreatic carcinoma. Hum Pathol 37:256–263PubMedCrossRefGoogle Scholar
  27. 27.
    Lee HS, Han J, Bai HJ et al (2009) Brain angiogenesis in developmental and pathological processes: regulation, molecular and cellular communication at the neurovascular interface. FEBS J 276:4622–4635PubMedCrossRefGoogle Scholar
  28. 28.
    Idbaih A, Carvalho Silva R, Criniere E et al (2008) Genomic changes in progression of low-grade gliomas. J Neurooncol 90:133–140PubMedCrossRefGoogle Scholar
  29. 29.
    van Agthoven T, Sieuwerts AM, Meijer-van Gelder ME et al (2009) Relevance of breast cancer antiestrogen resistance genes in human breast cancer progression and tamoxifen resistance. J Clin Oncol 27:542–549PubMedCrossRefGoogle Scholar
  30. 30.
    Blaschuk OW, Devemy E (2009) Cadherins as novel targets for anti-cancer therapy. Eur J Pharmacol 625:195–198PubMedCrossRefGoogle Scholar
  31. 31.
    Krishna K, Redies C (2009) Expression of cadherin superfamily genes in brain vascular development. J Cereb Blood Flow Metab 29:224–229CrossRefGoogle Scholar
  32. 32.
    Teodorczyk M, Martin-Villalba A (2009) Sensing invasion: cell surface receptors driving spreading of glioblastoma. J Cell Physiol 222:1–10CrossRefGoogle Scholar
  33. 33.
    Hayward SW, Wang Y, Cao M et al (2001) Malignant transformation in a nontumorigenic human prostatic epithelial cell line. Cancer Res 61:8135–8142PubMedGoogle Scholar
  34. 34.
    Orimo A, Gupta PB, Sgroi DC et al (2005) Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell 121:335–348PubMedCrossRefGoogle Scholar
  35. 35.
    Gonda TA, Varro A, Wang TC et al (2010) Molecular biology of cancer-associated fibroblasts: can these cells be targeted in anti-cancer therapy? Semin Cell Dev Biol 21:2–10PubMedCrossRefGoogle Scholar
  36. 36.
    Bauchet L, Mathieu-Daude H, Fabbro-Peray P et al (2010) Oncological patterns of care and outcome for 952 patients with newly diagnosed glioblastoma in 2004. Neuro OncolGoogle Scholar
  37. 37.
    Pietras K, Ostman A (2010) Hallmarks of cancer: interactions with the tumor stroma. Exp Cell Res 316:1324–1331PubMedCrossRefGoogle Scholar
  38. 38.
    Polyak K, Weinberg RA (2009) Transitions between epithelial and mesenchymal states: acquisition of malignant and stem cell traits. Nat Rev Cancer 9:265–273PubMedCrossRefGoogle Scholar
  39. 39.
    Phillips HS, Kharbanda S, Chen R et al (2006) Molecular subclasses of high-grade glioma predict prognosis, delineate a pattern of disease progression, and resemble stages in neurogenesis. Cancer Cell 9:157–173PubMedCrossRefGoogle Scholar
  40. 40.
    Tso CL, Shintaku P, Chen J et al (2006) Primary glioblastomas express mesenchymal stem-like properties. Mol Cancer Res 4:607–619PubMedCrossRefGoogle Scholar
  41. 41.
    Ricci-Vitiani L, Pallini R, Larocca LM et al (2008) Mesenchymal differentiation of glioblastoma stem cells. Cell Death Differ 15:1491–1498PubMedCrossRefGoogle Scholar
  42. 42.
    Gunther HS, Schmidt NO, Phillips HS et al (2008) Glioblastoma-derived stem cell-enriched cultures form distinct subgroups according to molecular and phenotypic criteria. Oncogene 27:2897–2909PubMedCrossRefGoogle Scholar
  43. 43.
    Rieske P, Golanska E, Zakrzewska M et al (2009) Arrested neural and advanced mesenchymal differentiation of glioblastoma cells-comparative study with neural progenitors. BMC Cancer 9:54PubMedCrossRefGoogle Scholar
  44. 44.
    Carro MS, Lim WK, Alvarez MJ et al (2010) The transcriptional network for mesenchymal transformation of brain tumours. Nature 463:318–325PubMedCrossRefGoogle Scholar
  45. 45.
    Nakamizo A, Marini F, Amano T et al (2005) Human bone marrow-derived mesenchymal stem cells in the treatment of gliomas. Cancer Res 65:3307–3318PubMedGoogle Scholar
  46. 46.
    Nakamura K, Ito Y, Kawano Y et al (2004) Antitumor effect of genetically engineered mesenchymal stem cells in a rat glioma model. Gene Ther 11:1155–1164PubMedCrossRefGoogle Scholar
  47. 47.
    Kang SG, Shinojima N, Hossain A et al (2010) Isolation and perivascular localization of mesenchymal stem cells from mouse brain. Neurosurgery 67:711–720PubMedCrossRefGoogle Scholar
  48. 48.
    Mishra PJ, Humeniuk R, Medina DJ et al (2008) Carcinoma-associated fibroblast-like differentiation of human mesenchymal stem cells. Cancer Res 68:4331–4339PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2011

Authors and Affiliations

  • Anne Clavreul
    • 1
    • 2
    • 3
  • Amandine Etcheverry
    • 4
    • 5
    • 6
    • 7
  • Agnès Chassevent
    • 8
  • Véronique Quillien
    • 5
    • 6
    • 9
  • Tony Avril
    • 9
  • Marie-Lise Jourdan
    • 10
    • 11
  • Sophie Michalak
    • 12
  • Patrick François
    • 13
  • Jean-Luc Carré
    • 14
  • Jean Mosser
    • 4
    • 5
    • 6
    • 7
  • The Grand Ouest Glioma Project Network
    • 15
  • Philippe Menei
    • 1
    • 2
    • 3
  1. 1.LUNAM Université, Ingénierie de la Vectorisation ParticulaireAngersFrance
  2. 2.INSERMAngersFrance
  3. 3.Département de NeurochirurgieCHU d’AngersAngersFrance
  4. 4.Plate-forme Génomique Santé Biogenouest®, IFR 140RennesFrance
  5. 5.CNRS, UMR 6061, Institut Génétique et Développement de RennesRennesFrance
  6. 6.Université Rennes 1, UEB, IFR 140, Faculté de MédecineRennesFrance
  7. 7.Service de Génétique Moléculaire et GénomiqueCHURennesFrance
  8. 8.Centre Régional de Lutte Contre le Cancer Paul PapinAngersFrance
  9. 9.Centre Eugène Marquis, Département de BiologieRennesFrance
  10. 10.INSERM U921ToursFrance
  11. 11.Laboratoire de CancérologieCHUToursFrance
  12. 12.Laboratoire Pathologie Cellulaire et Tissulaire, CHUAngersFrance
  13. 13.Service de Neurochirurgie, CHUToursFrance
  14. 14.Groupe Gliome BrestoisService de Biochimie et Biologie Moléculaire, UFR Médecine, CHUBrestFrance
  15. 15.Cancéropôle Grand OuestNantesFrance

Personalised recommendations