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Fig. 2 | BMC Cancer

Fig. 2

From: A way to understand idiopathic senescence and apoptosis in primary glioblastoma cells – possible approaches to circumvent these phenomena

Fig. 2

The comparison of NSC-like and classical monolayer conditions for culturing of primary glioblastoma cells. Both approaches did not enable to maintain long-term culture of GB cells and to avoid senescence/apoptosis; a-e Representative pictures showing results of the immunocytochemical analyses; a Early passage in monolayer serum conditions showing GFAP (+) GB cells; b Single GFAP (+), SA-β-Gal (+) GB cells from monolayer GB9 culture (intermediate passage) surrounded by cells negative for both these markers; c Early fresh GB9 neuropshere transferred onto poly-l-ornithine/laminin-coated dish, with the majority of GFAP (+) cells migrating from the neurosphere and only several αSMA (+) cells; d In late passage only large, flat and SA-β-Gal (+) cells with features of mitotic catastrophe maintained in culture; Inset boxes in B and D show higher magnifications of senescent (SA-β-Gal-positive) cells; e Intermediate passage of GB9 in NSC-like conditions – only GFAP (+) cells maintained in culture, but there is a lack of cells in mitosis; f Reduced proliferation rate of GB8 cells under NSC-like conditions vs. adherent monolayer culture; g-j The percentage of spontaneously senescent (as determined by means of SA-β-Gal assay), apoptotic (assessed using in vitro real-time caspase assay after 48 h of incubation with Caspase 3/7 synthetic reporter solution), proliferating (analyzed during 5 days incubation with BrdU), as well as other cells in various culture conditions and passages. The characteristics of cells in different culture conditions is depicted separately. The average percentage of cells with a particular feature was calculated by analyzing at least 200 cells per case from indicated passages of GB6-GB9 cultures. Error bars indicate SEM. k Statistical significance was calculated by paired Student’s t-test with p < 0.05 considered statistically significant

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