Additional file 1: Figure S1.

Phenotype, differentiation potential and immunosuppressive activity LA- and BM-MSC were cultured in 10% FBS, pHPL and tPRP, respectively. (A) Photos were taken at identical time points post-seeding to observe cell morphology and confluence. Representative pictures of one LA-MSC and one BM-MSC batch are depicted (all 100× magnification). (B) Adipogenic and osteogenic differentiation. Cells were induced with differentiation media (adipogenic induction and maintance medium for 3 and 4 days, respectively and osteogenic induction medium, all Lonza) for 3 weeks and then stained with oil red o and van Kossa as descried previously [11,13]. Representative results of one LA-MSC batch are depicted (100× magnification). (C) Immunosuppressive activity was assessed by coculturing LA-MSC (ratio 1:10) with allogeneic peripheral blood leukocytes labeled with carboxyfluorescein diacetate succinimidyl ester (5 μM, Vybrant CFDA-SE cell tracker kit, Invitrogen). Leukocyte proliferation was stimulated with phytohemagglutinin (2.5 μg/ml PHA-L, Roche Applied Science) and assessed by progressive halving of CFDA-SE fluorescence (red line – control with PHA; green – control without PHA). One representative experiment is depicted revealing similar dye retention and thus immunosuppressive activity of MSC in FBS, pHPL and tPRP (grey line – coculture MSC with PHA). Similar data were obtained applying BM-MSC.

Format: JPEG Size: 2.7MB Download file

Kinzebach et al. BMC Cell Biology 2013 14:48   doi:10.1186/1471-2121-14-48