Fig. 1

Isolation and proliferation capacity of bone-derived Nestin⁺ and Nestin⁻ cells. a Flow cytometry was used to isolate Nestin⁺ and Nestin⁻ cells in the gate of CD45⁻ Ter119⁻ CD31⁻ from the bone of Nestin-GFP transgenic mice. b Variations in morphology of the Nestin⁺ and Nestin⁻ cells were captured by microscopy examined at P3. Scale bar, 200 μm. c Growth curves of Nestin⁺ and Nestin⁻ cells as assessed by direct counting. Cells at P6 were seeded into a 12-well plate at 10,000 cells/well (triplicates), and the cells were then directly counted for a total of 6 days. d Colony-forming unit-fibroblast frequencies of Nestin⁺ and Nestin⁻ cells. Cells at P6 were seeded at a single cell per well into a 96-well plate. Colonies containing > 50 cells were counted under microscopic observation. The means ± SEMs of the results of three different experiments are shown. *p < 0.05; **p < 0.01; ***p < 0.001. e Phenotypic characterization of the cultured bone-derived Nestin⁺ and Nestin⁻ cells. Flow cytometry analysis of the presence of the cell surface markers Sca-1, c-kit, CD44, CD105, CD45, and CD11b on cultured bone-derived Nestin⁺ and Nestin⁻ cells