1. Redifferentiation of dedifferentiated human articular chondrocytes: comparison of 2D and 3D cultures 
M.M.J. Caron, P.J. Emans, M.M.E. Coolsen, L. Voss, D.A.M. Surtel, A. Cremers, L.W. van Rhijn, T.J.M. Welting 
Osteoarthritis and Cartilage 
Volume 20, Issue 10, Pages (October 2012)
DOI: /j.joca
Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

2. Fig. 1
RT-qPCR analysis of dedifferentiated HACs after redifferentiation in 2D and 3D culture systems. Dedifferentiated HACs (n=6 individuals) were redifferentiated in 2D monolayer or 3D alginate beads or pellet culture using identical medium conditions. Relative induction of gene expression was determined at day 7 in redifferentiation as compared to day 0. (A) Induction of mRNAs of the chondrogenic genes COL2A1, SOX9 and ACAN was determined. (B) Similar to (A) but here relative induction of hypertrophic/mineralization markers COL10A1, RUNX2, MMP13, VEGFA, OPN and ALP was determined. (C) Similar to (A) but now the fibrotic marker COL1A1 was determined. In all graphs dots represent the average value of three technical repeats per HAC isolate (n=6 individual donors). Statistics were calculated for these n=6 and error bars represent mean±95% confidence interval. Statistical significant P-values (P<0.05) are shown in bold/italic.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

3. Fig. 2
Immunoblot analysis of dedifferentiated HACs after redifferentiation in 2D and 3D culture systems. HACs (n=6 individuals) were redifferentiated in monolayer, alginate beads or pellet culture. Protein expression of a subset of the genes from Fig. 1 (COL2A1, COL10A1, COL1A1, SOX9, RUNX2) was determined at day 7 in redifferentiation as compared to day 0. (A) Protein expression of HACs redifferentiated in 2D monolayer culture. (B) Protein expression of HACs redifferentiated in 3D alginate beads. (C) Protein expression of HACs redifferentiated in 3D pellet cultures. α-Tubulin was used as housekeeper. Quantifications of ECL signals (corrected for the complementary α-Tubulin signals and relatively to t=0) are depicted on top of all immunoblots. In immunoblots COL2A1 runs at 120kDa; COL10A1 at 180kDa; COL1A1 at 130kDa; SOX9 at 60kDa; RUNX2 at 66kDa (lower band). (D) Graphs of immunoblot quantifications from (A) to (C). Each dot represents the determined value for each of the individual HAC isolates (n=6) and error bars represent mean±95% confidence interval. Statistical significant P-values (P<0.05) are shown in bold/italic.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

4. Fig. 3
GAG content and DNA induction in redifferentiated HACs. Dedifferentiated HACs (n=6 individuals) were redifferentiated in monolayer, alginate beads or pellet culture, using identical medium conditions. (A) Total GAG content was determined in day 0 and day 7 samples. To correct for cells numbers, GAG content was normalized to total DNA. (B) The DNA content used in (A) to correct GAG content is shown to determine cell proliferation. DNA induction was calculated relative to DNA content in day 0 samples. In the graphs dots represent the average value of three technical repeats per HAC isolate (n=6 individual donors). Statistics were calculated for these n=6 and error bars represent mean±95% confidence interval. Statistical significant P-values (P<0.05) are shown in bold/italic.
Osteoarthritis and Cartilage  , DOI: ( /j.joca )
Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions