Abstract
Glioblastomas are brain tumors derived from astrocytes or oligodendrocytes. These tumors have a heterogeneous structure composed of a necrotic and vascularized center and an invasive periphery. The rapid growth of glioblastoma and its ability to invade surrounding tissues make this cancer difficult to treat. The median survival of patients afflicted with the disease ranges from 12 to 15 months. The standard treatment, based on the Stupp protocol, consisting of a resection of the tumor mass and adjuvant temozolomide treatment and/or radiotherapy, allows delaying recurrence for only a few months. Since tumor cell invasion is the central element in tumor recurrence, it is important to understand the mechanisms of tumor invasion and the interactions between tumor cells and their microenvironment. To this aim, suitable models have been developed and are constantly updated. In this article, we describe in detail experimental procedures based on the spheroid technology which allow mechanistical studies related to proliferation, survival, migration, and invasion.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Kotliarova S, Fine HA (2012) SnapShot: glioblastoma multiforme. Cancer Cell 21(5):710–710. e711. https://doi.org/10.1016/j.ccr.2012.04.031
Stupp R, Hegi ME, Gilbert MR, Chakravarti A (2007) Chemoradiotherapy in malignant glioma: standard of care and future directions. J Clin Oncol Off J Am Soc Clin Oncol 25(26):4127–4136. https://doi.org/10.1200/JCO.2007.11.8554
Gatson NN, Chiocca EA, Kaur B (2012) Anti-angiogenic gene therapy in the treatment of malignant gliomas. Neurosci Lett 527(2):62–70. https://doi.org/10.1016/j.neulet.2012.08.001
Weller M, Yung WK (2013) Angiogenesis inhibition for glioblastoma at the edge: beyond AVAGlio and RTOG 0825. Neuro-Oncology 15(8):971. https://doi.org/10.1093/neuonc/not106
Patel AP, Tirosh I, Trombetta JJ, Shalek AK, Gillespie SM, Wakimoto H, Cahill DP, Nahed BV, Curry WT, Martuza RL, Louis DN, Rozenblatt-Rosen O, Suva ML, Regev A, Bernstein BE (2014) Single-cell RNA-seq highlights intratumoral heterogeneity in primary glioblastoma. Science 344(6190):1396–1401. https://doi.org/10.1126/science.1254257
Lenting K, Verhaak R, Ter Laan M, Wesseling P, Leenders W (2017) Glioma: experimental models and reality. Acta Neuropathol 133(2):263–282. https://doi.org/10.1007/s00401-017-1671-4
Torsvik A, Stieber D, Enger PO, Golebiewska A, Molven A, Svendsen A, Westermark B, Niclou SP, Olsen TK, Chekenya Enger M, Bjerkvig R (2014) U-251 revisited: genetic drift and phenotypic consequences of long-term cultures of glioblastoma cells. Cancer Med 3(4):812–824. https://doi.org/10.1002/cam4.219
Newcomb EW, Lukyanov Y, Alonso-Basanta M, Esencay M, Smirnova I, Schnee T, Shao Y, Devitt ML, Zagzag D, McBride W, Formenti SC (2008) Antiangiogenic effects of noscapine enhance radioresponse for GL261 tumors. Int J Radiat Oncol Biol Phys 71(5):1477–1484. https://doi.org/10.1016/j.ijrobp.2008.04.020
Bougnaud S, Golebiewska A, Oudin A, Keunen O, Harter PN, Mader L, Azuaje F, Fritah S, Stieber D, Kaoma T, Vallar L, Brons NH, Daubon T, Miletic H, Sundstrom T, Herold-Mende C, Mittelbronn M, Bjerkvig R, Niclou SP (2016) Molecular crosstalk between tumour and brain parenchyma instructs histopathological features in glioblastoma. Oncotarget 7(22):31955–31971. https://doi.org/10.18632/oncotarget.7454
Wang J, Miletic H, Sakariassen PO, Huszthy PC, Jacobsen H, Brekka N, Li X, Zhao P, Mork S, Chekenya M, Bjerkvig R, Enger PO (2009) A reproducible brain tumour model established from human glioblastoma biopsies. BMC Cancer 9:465. https://doi.org/10.1186/1471-2407-9-465
Huszthy PC, Daphu I, Niclou SP, Stieber D, Nigro JM, Sakariassen PO, Miletic H, Thorsen F, Bjerkvig R (2012) In vivo models of primary brain tumors: pitfalls and perspectives. Neuro-Oncology 14(8):979–993. https://doi.org/10.1093/neuonc/nos135
Kim SS, Pirollo KF, Chang EH (2015) Isolation and culturing of glioma cancer stem cells. Curr Protoc Cell Biol 67:23.10.1–21.10.10. https://doi.org/10.1002/0471143030.cb2310s67
Boye K, Pujol N, Isabel DA, Chen YP, Daubon T, Lee YZ, Dedieu S, Constantin M, Bello L, Rossi M, Bjerkvig R, Sue SC, Bikfalvi A, Billottet C (2017) The role of CXCR3/LRP1 cross-talk in the invasion of primary brain tumors. Nat Commun 8(1):1571. https://doi.org/10.1038/s41467-017-01686-y
Daubon T, Leon C, Clarke K, Andrique L, Salabert L, Darbo E, Pineau R, Guerit S, Maitre M, Dedieu S, Jeanne A, Bailly S, Feige JJ, Miletic H, Rossi M, Bello L, Falciani F, Bjerkvig R, Bikfalvi A (2019) Deciphering the complex role of thrombospondin-1 in glioblastoma development. Nat Commun 10(1):1146. https://doi.org/10.1038/s41467-019-08480-y
Zimmerman NP, Roy I, Hauser AD, Wilson JM, Williams CL, Dwinell MB (2015) Cyclic AMP regulates the migration and invasion potential of human pancreatic cancer cells. Mol Carcinog 54(3):203–215. https://doi.org/10.1002/mc.22091
Allen E, Jabouille A, Rivera LB, Lodewijckx I, Missiaen R, Steri V, Feyen K, Tawney J, Hanahan D, Michael IP, Bergers G (2017) Combined antiangiogenic and anti-PD-L1 therapy stimulates tumor immunity through HEV formation. Sci Transl Med 9(385). https://doi.org/10.1126/scitranslmed.aak9679
Huse JT, Holland EC (2009) Genetically engineered mouse models of brain cancer and the promise of preclinical testing. Brain Pathol 19(1):132–143. https://doi.org/10.1111/j.1750-3639.2008.00234.x
Acknowledgments
Joris Guyon is a recipient of Toulouse Hospital scholarship, and Tiffanie Chouleur is a PhD student supported by IDEX Bordeaux-McGill University. Andreas Bikfalvi got financial support from Ligue Contre le Cancer, ARC, ANR, and TRANSCAN-ERA-NET. Thomas Daubon is supported by ARC and ERA-NET grants.
Author Contributions
JG, TC, AB, and TD wrote the manuscript. JG designed all Figs. TD and AB discussed the manuscript.
Author information
Authors and Affiliations
Corresponding authors
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2021 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Guyon, J., Chouleur, T., Bikfalvi, A., Daubon, T. (2021). Glioblastoma Patient-Derived Cell Lines: Generation of Nonadherent Cellular Models from Brain Tumors. In: Seano, G. (eds) Brain Tumors. Neuromethods, vol 158. Springer, New York, NY. https://doi.org/10.1007/978-1-0716-0856-2_5
Download citation
DOI: https://doi.org/10.1007/978-1-0716-0856-2_5
Published:
Publisher Name: Springer, New York, NY
Print ISBN: 978-1-0716-0855-5
Online ISBN: 978-1-0716-0856-2
eBook Packages: Springer Protocols