Figure 3
Differential expression of chondrogenic and osteogenic markers for UCM-MSCs and UCB-MSCs after chondrogenic differentiation in type I/III collagen scaffolds. UCM-MSCs and UCB-MSCs were cultured in type I/III collagen scaffolds and normoxia for 28 days in incomplete chondrogenic medium (ICM) or ICM supplemented with BMP-2 (50 ng/ml) and TGF-ß1 (10 ng/ml) (BT). Relative mRNA amounts encoding chondrogenic markers (A) (Col2a1, Acan and Sox9), hypertrophic markers (C) (Col10a1 and Mmp13) and osteogenic markers (D) (Col1a1, Alpl, Spp1, Htra1, Runx2 and Bglap) were determined by RTqPCR. The functional Col2a1/Col1a1 ratio was presented in (B) mRNA extracts obtained from equine articular chondrocytes (eAC) released from cartilage after overnight enzymatic digestion and equine P0 osteoblasts (eO) were used as controls. Monolayer cultured MSCs (2D) were used as undifferentiated cells control. All results for each MSCs source were normalized versus untreated MSCs incubated during 1 h in type I/III collagen scaffolds at 37 °C and presented as the relative expression of each gene. Box-plots represent five independent experiments for each MSCs source. Statistically significant differences were determined using the Mann-Whitney test. *: 2D versus other conditions (*p < 0.05, **p < 0.01). §: ICM versus other conditions (§p < 0.05, §§p < 0.01). (E) Steady-state levels of mRNA expression in UCB-MSCs and UCM-MSCs after 28 days of chondrogenic differentiation in BT condition were compared by RTqPCR. All results were normalized versus undifferentiated sample of one UCM-MSCs sample, and presented as the relative expression of each gene. Box-plots represent five independent experiments and statistically significant differences were determined using the Mann Whitney test by comparison of UCM-MSCs versus UCB-MSCs *:(*p < 0.05, **p < 0.01).
