Fig. 2

Culture conditions can interfere with the fate of GBM CD105⁺ cells. A Schematic representation of media conditions for CD105⁺ primary cell culture. B Primary cell culture in SC and SFC under passage 3 (P3). CD105⁺ cells and neural stem-like cells were identified by staining of CD105 (red) and SOX2 (green), respectively. C Flow cytometry analysis of CD105⁺ cells in different media conditions. Flow cytometry data show the GBM CD105 subpopulation obtained from GBM primary cells using the phycoerythrin (PE) channel. ***P < 0.001. D Immunostaining of CD105⁺ cell with cell type markers. Double immunostaining of CD105 (red) and cellular markers (green) on sorted CD105⁺ cells (under P3). E, F Effect of serum/ serum-free culture conditions on the differentiation of GBM primary cell lines. SFC cultured SOX2⁺ cells differentiated into DMEM/F12 media supplemented with 10% FBS for 14 days (E). Immunostaining of CD105 (red) and SOX2 (green) shown on differentiated cells (DAPI blue). Bar graph showing cell viability of 5 GBM SOX2⁺ cell lines differentiated into SC and SFC for 14 days. An exceptional SOX2⁺ cell line, GBM B17, differentiated in SC with different serum concentrations, showed a higher percentage of CD105⁺ cells by flow cytometry data. F Sorted CD105⁺ cells differentiated in SFC supplemented with b-FGF for 14 days. Differentiated cells were stained with CD105 (red) and SOX2 (green). Bar graphs showing quantification of cell viability of (left lower) and CD105⁺ cell subpopulation (right lower). 5 CD105⁺ cell lines were differentiated in SFC for 14 days. Graph shows CD105⁺ cell lines tolerating SFC but losing CD105 marker positivity. *P < 0.05 **P < 0.01 ***P < 0.001