Table 1.
Primers used for qPCR analysis.
Fig 1.
Morphology of hWJ-MSCs with a typical fibroblast-like morphology.
(A) Phase contrast images of hWJ-MSCs expanded from Wharton’s jelly tissue (arrow) and (B) hWJ-MSCs at 80% confluences. Scale bar = 10 μm.
Fig 2.
(A) Immunophenotype of MSCs, immunofluorescent micrographs staining expression of MSC markers (CD73, 90, and 105), Nuclei were counterstains with DAPI (blue). Cells were negative for hematopoietic marker (CD34). Scale bar = 20 μm. (B) Differentiation of hWJ-MSCs to mesodermal linage cells. The cells were induced to undergo adipogenic, osteogenic, and condrogenic differentiation.
Fig 3.
The toxicity effect of LiCl and SB216763 on hWJ-MSC viability.
hWJ-MSCs were cultured with 0–20 mM LiCl (A), 0–5 μM SB216763 (B), or 0–0.5% DMSO (C), for 72 hrs in 96-well plate. Then, the viability was detected by MTT assay. **DMSO without chemical was use as vehicle control. **Data were exposed as mean ± SD. *P<0.05.
Fig 4.
Accumulation of GAGs was stained by alcian blue.
(A-D) Photographs of monolayer expanded cells cultured for 2 weeks. Scale bar = 10 μm. (E-H) Pellets culture at 4 weeks after differentiation. Scale bar = 20 μm. (I) Morphology of pellet culture at 4 weeks after differentiation.
Fig 5.
Immunofluorescent staining of cartilage specific type collagen.
(A) immunofluorescent staining for collagen type II in monolayer expanded cultured on 3 weeks after differentiation. Scale bar = 20 μm. (B) Collagen type II and X expressions in pellet experiment on 4 weeks after differentiation. Scale bar = 20 μm.
Fig 6.
The collagen type II protein after 4 weeks of inductions examined by western blot analysis.
β-actin was used as an internal control.
Fig 7.
qPCR analysis for chondrogenic gene expressions after 4 weeks of inductions.
(A) Col2a1, (B) ACAN, (C) Sox9 and (D) β-catenin. Gene expression was normalized to coresponding GAPDH and calculated by relative expression compared to control cells. The experiments were perfromed three times. **Data were expressed as mean±SD, *P<0.05.
Fig 8.
Expression level of hypertrophic marker genes (A) Col10a1, and (B) Runx2 was quantified by qPCR after 2, 3, and 4 weeks of inductions.
Gene expression was normalized to coresponding GAPDH and calculated by relative expression compared to control cells. Data were expressed as mean±SD, The expperiments were perfromed three times.