Phenotypic and functional characterization of primary murine MSCs.

<p>A) Immunophenotype of bone marrow-derived MSCs isolated from C57BL/6 and DBA/1 mice. Cells were stained with antibodies against the indicated antigens and analysed by flow cytometry. Representative histograms are shown in black and the respective isotype controls are shown as grey line and are representative of three experiments. B) Differentiation potential of murine MSCs. The differentiation in the 3 lineages is shown by up-regulation of specific markers by RT-qPCR and specific staining. Osteogenesis is characterized by the expression of osteocalcin (OC), alkaline phosphatase (AP) and Alizarin Red S positive staining in differentiation medium (lower panel) versus proliferative medium (upper panel). Adipogenesis is characterized by the expression of peroxysome proliferator-activated receptor (PPAR)-γ; fatty acid binding protein 4 (aP2) and Oil Red O positive staining in differentiation medium (lower panel) versus proliferative medium (upper panel). Chondrogenesis is characterized by the expression of collagen II (Col II) and aggrecan (Agg) and anti-aggrecan positive staining on pellet sections (lower panel) versus unstained control section (upper panel). Results are representative of three independent experiments. C) Immunosuppressive potential of murine C57BL/6 (left panel) and DBA/1 (right panel) MSCs. Allogeneic splenocytes were incubated for three days with ConA without or with various ratios of MSCs. Results are expressed as the percentage of ConA-induced proliferation which was assigned the value of 100%± SEM (n = 3; ***: p<0.005).