|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Identification of cancer stem cells from human glioblastomas: growth and differentiation capabilities and CD133/prominin-1 expression
Federica Gambelli*, Federica Sasdelli*, Ivana Manini*, Carla Gambarana†, Giuseppe Oliveri‡, Clelia Miracco§ and Vincenzo Sorrentino*1
*Molecular Medicine Section, Department of Neuroscience and Center for Stem Cell Research, University of Siena, Siena, Italy, †Pharmacology Unit, Department of Neuroscience, University of Siena, Siena, Italy, ‡Department of Neurosurgery, Santa Maria alle Scotte Hospital, Siena, Italy, and §Section of Pathological Anatomy, Department of Human Pathology and Oncology, University of Siena, Siena, Italy
CD133 can be a marker of tumorigenic CSCs (cancer stem cells) in human GBM (glioblastoma multiforme), although tumorigenic CD133-negative CSCs have been also isolated. Additional evidence indicates that CSCs from GBM exhibit different phenotypes, with increasing interest in the potential significance of the different CSCs with respect to diagnosis, prognosis and the development of novel targets for treatment. We have analysed the expression of CD133 in freshly isolated cells from 15 human GBM specimens. Only 4 of them contained cells positive for AC133 by FACS analysis, and all of them yielded distinct CSC lines, whereas only 6 CSC lines were obtained from the other 11 GBMs. Of these 10 CSCs lines, we further characterized 6 CSC lines. Three CSCs grew as fast-growing neurospheres with higher clonogenic ability, whereas the remaining 3 grew as slow-growing semi-adherent spheres of lower clonogenicity. In addition, the former CSC lines displayed better differentiation capabilities than the latter ones. PCR and Western blot analysis showed that all 6 GBM CSC lines expressed CD133/prominin-1, suggesting that cells negative by FACS analysis may actually represent cells expressing low levels of CD133 undetected by FACS. Nevertheless, all the 6 CSC lines were tumorigenic in nude mice. In conclusion, CSCs from human primary GBMs show different phenotypes and variable levels of CD133 expression, but these parameters did not directly correlate with the tumorigenic potential.
Key words: brain cancer stem cells, CD133, glioblastoma, tumour stem cell marker
Abbreviations: BSA, bovine serum albumin, CSC, cancer stem cell, CXCR4, CXC chemokine receptor 4, DAPI, 4′,6-diamidino-2-phenylindole, DMEM, Dulbecco's modified Eagle's medium, GBM, glioblastoma multiforme, GalC, galactocerebroside C, GBM, glioblastoma multiforme, GFAP, glial fibrillary acidic protein, LDA, limiting dilution assay, mAb, monoclonal antibody, MSC, mesenchymal stem cell, NF-H, neurofilament heavy polypeptide, NSC, neural stem cell, PE, phycoerythrin, RT–PCR, reverse transcription–PCR, SOX-2, SRY-related HMG-box gene 2, TBS-T, Tris-buffered saline with 0.1% Tween 20
1To whom correspondence should be addressed (email email@example.com).
Received 8 January 2011/19 April 2011; accepted 15 September 2011
Published as Cell Biology International Immediate Publication 15 September 2011, doi:10.1042/CBI20110013
© The Author(s) Journal compilation © 2012 Portland Press Limited