MAPCs derived from sorted GFP-positive BM mononuclear cells were cultured for more than 50 cell doublings, then induced to differentiate into endothelial cells (mesodermal lineage), hepatocytes (endodermal lineage), or neurons (ectodermal lineage). (A,D,G) GFP-positive MAPCs treated with VEGF (A-C; notice vascular tubelike structures), FGF-4/HGF (D-F), or bFGF (G-I) for 14 days. Cultures were stained with anti-VWF labeled with Cy3 (B), anti-albumin-Cy3 (E), or anti-NF200-Cy3 (H). (C,F,I) An overlay of Cy3 staining with green fluorescence and DAPI. (A,C,D,F, G,I) 100% of these cells were eGFP positive. (J) Cytogentic analysis of selected clones revealed a diploid karyotype. (K) BM from secondary recipients showed trilineage hematopoietic differentiation (% GFP-positive cells determined by FACS). (L) MAPCs from secondary recipients contained GFP-positive cells. Inset: Binucleated cells. Bar equals 200 μm.