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

Abstract

BackgroundPatient-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 stem-like 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 methodsTo address this question, we used two heterogeneous glioblastoma GSC lines (NCH421k and NCH644) that lack EGFR amplification.ResultsAlthough 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).ConclusionsWe 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.

Differentially expressed proteins in glioblastoma multiforme identified with a nanobody-based anti-proteome approach and confirmed by OncoFinder as possible tumor-class predictive biomarker candidates

“…Protein extracts used for phage particle enrichment and during panning were obtained from two commercially available GBM stem-like cell lines: NCH644 and NCH421K cells (Cell Line Service, Eppelheim, Germany) [ 49 , 50 ]. Cells were grown in complete medium, as neurobasal medium (Gibco) plus 20 μM L-glutamine, 100 U/mL penicillin, 100 μg/mL streptomycin, 2% B-27 supplement, 20 ng/mL basic fibroblast growth factor, 20 ng/mL epidermal growth factor, and 1 U/mL heparin.…”

Targeting APLN/APLNR Improves Antiangiogenic Efficiency and Blunts Proinvasive Side Effects of VEGFA/VEGFR2 Blockade in Glioblastoma

“…We also orthotopically implanted mice with a second wellestablished human primary GSC model (NCH644 cells (20)) corresponding to the proneural subtype (36). These cells produced a GBM with a VLD of 1,168 mm/mm 3 in wild-type mice ( Supplementary Fig.…”

Expression and Significance of IL-6 and bFGF in Human Tongue Squamous Cell Carcinoma

“…Information available at present suggests that the biological functions of bFGF are extensive, and bFGF is very important in the promotion of angiopoiesis, wound healing and tissue repair, tissue regeneration, and the growth and development of nervous tissue [14] [15] [16]. The expression of bFGF in squamous cell carcinoma is significantly higher than that in normal groups and benign tumor and the expression rate in high differentiation groups is significantly higher than that in the other two.…”