Figure 2: Differentiation assay, single-cell cloning and telomerase activity assay of SM-MHC − cells derived from rat carotid arterial tunica media.

(a–f) Staining of differentiated cell derived from SM-MHC⁻ cells: Schwann cells for GFAP (a), neurons for TUJ1 (b), SMCs for SM-MHC (c), chondrocytes for aggrecan using alcian blue (d), adipocytes for lipid droplets using oil red (e) and osteoblasts for calcified matrix using alizarin red (f). Scale bars of (a–c) are 50 μm. Scale bars of (d–f) are 100 μm. (g) Schematic illustration of single-cell cloning with maintenance media. (h–i) Immunostaining of cloned MVSCs for Sox10 and Sox17. (j) Telomerase activity assay of MVSCs and the tissues from which MVSCs were isolated. The data was shown as average±s.d. (n=3). White bars indicate tissues, and black bars indicate isolated MVSCs. * indicates significant difference between MVSCs and the tissue from which the cells were derived using Student's t-test (P<0.05). † indicates significant difference between inferior vena cava and other blood vessels using Holm's t-test (P<0.05). AO, aorta, CA, carotid artery, JV, jugular vein, AA, abdominal artery, IVC, inferior vena cava, FA, femoral artery, FV, femoral vein. (k) DNA microarray analysis of MVSCs derived from rat carotid arteries and jugular veins (n=3).