<abstract xmlns="http://www.w3.org/1999/xhtml"> Background. Patient-derived glioblastoma (GBM) stem-like cells (GSCs) represent a valuable model for basic and therapeutic research. GSCs are usually propagated in serum-free Neural Basal medium supplemented with bFGF and EGF. Yet, the exact influence of these growth factors on GSCs is still unclear. Recently it was suggested that GBM stemlike cells with amplified EGFR should be cultured in stem cell medium without EGF, as the presence of EGF induced rapid loss of EGFR amplification. However, patient biopsies are usually taken into culture before their genomic profiles are defined. Thus, an important question remains whether GBM cells without EGFR amplification also can be cultured in stem cell medium without EGF. Meterials and methods. To address this question, we used two heterogeneous glioblastoma GSC lines (NCH421k and NCH644) that lack EGFR amplification. Results. Although both cell lines showed very low EGFR expression under standard growth conditions, bFGF stimulation induced higher expression of EGFR in NCH644. In both cell lines, expression of the stem cell markers nestin and CD133 was higher upon stimulation with bFGF compared to EGF. Importantly, bFGF stimulated the growth of both cell lines, whereas EGF had no effect. We verified that the growth stimulation by bFGF was either mediated by proliferation (NCH421k) or resistance to apoptosis (NCH644). Conclusions. We demonstrate that GSC cultures without EGFR amplification can be maintained and expanded with bFGF, while the addition of EGF has no significant effect and therefore can be omitted.</abstract>

Background. Patient-derived glioblastoma (GBM) stem-like cells (GSCs) represent a valuable model for basic and therapeutic research. GSCs are usually propagated in serum-free Neural Basal medium supplemented with bFGF and EGF. Yet, the exact influence of these growth factors on GSCs is still unclear. Recently it was suggested that GBM stemlike cells with amplified EGFR should be cultured in stem cell medium without EGF, as the presence of EGF induced rapid loss of EGFR amplification. However, patient biopsies are usually taken into culture before their genomic profiles are defined. Thus, an important question remains whether GBM cells without EGFR amplification also can be cultured in stem cell medium without EGF. Meterials and methods. To address this question, we used two heterogeneous glioblastoma GSC lines (NCH421k and NCH644) that lack EGFR amplification. Results. Although both cell lines showed very low EGFR expression under standard growth conditions, bFGF stimulation induced higher expression of EGFR in NCH644. In both cell lines, expression of the stem cell markers nestin and CD133 was higher upon stimulation with bFGF compared to EGF. Importantly, bFGF stimulated the growth of both cell lines, whereas EGF had no effect. We verified that the growth stimulation by bFGF was either mediated by proliferation (NCH421k) or resistance to apoptosis (NCH644). Conclusions. We demonstrate that GSC cultures without EGFR amplification can be maintained and expanded with bFGF, while the addition of EGF has no significant effect and therefore can be omitted.

Expansive growth of two glioblastoma stem-like cell lines is mediated by bFGF and not by EGF

Department of Genetic Toxicology and Cancer Biology, National Institute of Biology, Ljubljana, Slovenia

Department of Biomedicine, University of Bergen, Bergen, Norway

German Cancer Research Centre, Division of Molecular Genetics, Heidelberg, Germany

Division of Neurosurgical Research, Department of Neurosurgery, University of Heidelberg, Heidelberg, Germany

NorLux Neuro-Oncology Laboratory, Centre de Recherche Public de la Santé, Luxembourg

Blood Transfusion Centre of Slovenia, Ljubljana, Slovenia

Department of Biochemistry, Faculty of Chemistry and Chemical Engineering, University of Ljubljana, Ljubljana, Slovenia

Department of Pathology, Haukeland University Hospital, Bergen, Norway

Radiology and Oncology

Background. Patient-derived glioblastoma (GBM) stem-like cells (GSCs) represent a valuable model for basic and therapeutic research. GSCs are usually...

Expansive growth of two glioblastoma stem-like cell lines is mediated by bFGF and not by EGF

Publié en ligne: 08 oct. 2013

Pages: 330 - 337

DOI: https://doi.org/10.2478/raon-2013-0063

Mots clés
> glioblastoma, bFGF, EGF, EGFR, stem cell cultures

This content is open access.

Expansive growth of two glioblastoma stem-like cell lines is mediated by bFGF and not by EGF

Publié en ligne: 08 oct. 2013

Pages: 330 - 337

DOI: https://doi.org/10.2478/raon-2013-0063

Mots clés> glioblastoma, bFGF, EGF, EGFR, stem cell cultures

This content is open access.

Mots clés
> glioblastoma, bFGF, EGF, EGFR, stem cell cultures