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. 2014 Mar 6;3(4):433–447. doi: 10.5966/sctm.2013-0138

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Figure 5. — c-iPSCs form articular cartilage matrix in three-dimensional (3D) pellet mass cultures. (A): Histological examination of 3D pellet mass cultures stained with Alcian blue van Gieson showing chondrocytes from donor A2 before reprogramming at 2 weeks of differentiation as control and A2B c-iPSCs at 2 weeks and 5 weeks of differentiation. Blue color stains proteoglycans in the extracellular matrix. Scale bar = 100 μm. (B): Histological sections of the differentiated A2B c-iPSCs in pellet mass culture, stained with 4′,6-diamidino-2-phenylindole (DAPI) for cell nuclei and collagen II antibody at 2 weeks and 5 weeks in culture. Sections of pellet mass culture of chondrocytes from donor A2 are shown as control. Scale bar = 200 μm. Staining shows increased collagen II content in the pellets with time in culture. (C): Relative gene expression patterns of chondrogenic markers ACAN (Ci), SOX9 (Cii), COL2A1 type A (Ciii), COL2A1 type B (Civ); pluripotency markers OCT4 (Cv) and NANOG (Cvi); and hypertrophic marker COL10A1 (Cvii). From left to right, the donor chondrocytes in 3D pellet mass cultures before reprogramming, donor chondrocytes in monolayer just before reprogramming, A2B c-iPSCs, prechondrogenic cells, A2B c-iPSCs differentiated for 2 weeks, and A2B c-iPSCs differentiated for 5 weeks. Results shown as mean (n = 3; chondrocyte monolayer, n = 1) with error bars showing standard deviation. Expressions normalized to highest expression for each separate gene. ∗, p < .05, ∗∗, p < .01, ∗∗∗, p < .001. Abbreviations: 2w, 2 weeks; 5w, 5 weeks; c-iPS, chondrocyte-derived induced pluripotent stem cells; hEL, human diploid embryonic lung fibroblast cells; Rel., relative.

Figure 5. — c-iPSCs form articular cartilage matrix in three-dimensional (3D) pellet mass cultures. (A): Histological examination of 3D pellet mass cultures stained with Alcian blue van Gieson showing chondrocytes from donor A2 before reprogramming at 2 weeks of differentiation as control and A2B c-iPSCs at 2 weeks and 5 weeks of differentiation. Blue color stains proteoglycans in the extracellular matrix. Scale bar = 100 μm. (B): Histological sections of the differentiated A2B c-iPSCs in pellet mass culture, stained with 4′,6-diamidino-2-phenylindole (DAPI) for cell nuclei and collagen II antibody at 2 weeks and 5 weeks in culture. Sections of pellet mass culture of chondrocytes from donor A2 are shown as control. Scale bar = 200 μm. Staining shows increased collagen II content in the pellets with time in culture. (C): Relative gene expression patterns of chondrogenic markers ACAN (Ci), SOX9 (Cii), COL2A1 type A (Ciii), COL2A1 type B (Civ); pluripotency markers OCT4 (Cv) and NANOG (Cvi); and hypertrophic marker COL10A1 (Cvii). From left to right, the donor chondrocytes in 3D pellet mass cultures before reprogramming, donor chondrocytes in monolayer just before reprogramming, A2B c-iPSCs, prechondrogenic cells, A2B c-iPSCs differentiated for 2 weeks, and A2B c-iPSCs differentiated for 5 weeks. Results shown as mean (n = 3; chondrocyte monolayer, n = 1) with error bars showing standard deviation. Expressions normalized to highest expression for each separate gene. ∗, p < .05, ∗∗, p < .01, ∗∗∗, p < .001. Abbreviations: 2w, 2 weeks; 5w, 5 weeks; c-iPS, chondrocyte-derived induced pluripotent stem cells; hEL, human diploid embryonic lung fibroblast cells; Rel., relative.