Fig. 3

Multilineage differentiation of CB-MSC. Multilineage ability was determined in P4 LL-CBMSC. a-f Panels display cells which have been induced to differentiate in vitro toward osteogenic (a-b), adipogenic (c-d), and chondrogenic (e-f) lineages. Osteogenic and adipogenic differentiation were assessed after 21 days of induction using von Kossa and Oil Red O staining, respectively; ×10 magnification. Chondrogenesis was evaluated by Alcian Blue staining at day 28 of induction; cells were counterstained with Nuclear Fast Red solution; ×20 magnification. For each staining, undifferentiated controls are also displayed on the left (panels a-c-e). g Quantitative RT-PCR analysis of osteogenic markers RUNX2 and ALP (g-h), adipogenic markers PPARG and FABP4 (i-j), and chondrogenic markers SOX9 and COLXA1 (k-l) in cells cultured under the respective lineage induction conditions. Results are presented as the fold change in mRNA expression in respect to TBP as representative reference gene and to the undifferentiated control. The mean values from three independent experiments done in triplicate are shown. The differences were computed by paired t test or Wilcoxon matched pairs test as appropriate, p values: ^** p < 0.01. Abbreviations: NS not significant