Fig. 3

CD105⁺ cell stemness and tumorigenicity assay. A Schematic of intracranial transplantation for CD105⁺ cells. Sorted primary CD105⁺ cell lines (n=5) cultured in 2D and 3D conditions. 250,000 cells were injected into NSG mouse brains. Brain tissue was harvested immediately following the loss of each animal. MSCs cell line from a healthy donor and U87 cell line was used as controls (n=5 in each group). B Sphere formation assay of GBM CD105⁺ cells. CD105⁺ cells were cultured in 3D conditions for 14 days and tumorspheres captured under the brightfield or fluorescence microscopy. IF double staining shows CD105 double stained together with SOX2, Nestin and Ki67. C Comparison of the cell viability of CD105⁺ cells between 2D and 3D cultured conditions. 1000 of 2D or 3D CD105⁺ cells cultured for 24h. Cell viability assay based on PrestoBlue fluorescence intensity of each CD105⁺ cell line. *** P < 0.001, *P < 0.05. D Kaplan–Meier survival curves comparing 2D or 3D cultured CD105⁺ cells xenografts. E Scans of mouse brain tumor sections. F Brain sections of xenografted mice transplanted with 2D or 3D cultured CD105⁺ cells stained with CD105 (red) and Hu (green) and DAPI (blue)