M. M. J. Caron, P. J. Emans, M. M. E. Coolsen, L. Voss, D. A. M

Slides:

Advertisements

Similar presentations

M. M. J. Caron, P. J. Emans, M. M. E. Coolsen, L. Voss, D. A. M

Advertisements

Expression pattern differences between osteoarthritic chondrocytes and mesenchymal stem cells during chondrogenic differentiation P. Bernstein, C. Sticht,

Chondro-protective effects of low intensity pulsed ultrasound

Bone morphogenetic protein (BMP)-2 enhances the expression of type II collagen and aggrecan in chondrocytes embedded in alginate beads Tatiana Gründer,

Time-lapse observation of the dedifferentiation process in mouse chondrocytes using chondrocyte-specific reporters Y. Minegishi, K. Hosokawa, N. Tsumaki

Muscle cell-derived factors inhibit inflammatory stimuli-induced damage in hMSC- derived chondrocytes R.S. Rainbow, H. Kwon, A.T. Foote, R.C. Preda, D.L.

GDF5 reduces MMP13 expression in human chondrocytes via DKK1 mediated canonical Wnt signaling inhibition L. Enochson, J. Stenberg, M. Brittberg, A. Lindahl

Regulation of mesenchymal stem cell chondrogenesis by glucose through protein kinase C/transforming growth factor signaling T.-L. Tsai, P.A. Manner,

Insulin-like growth factor-1 boosts the developing process of condylar hyperplasia by stimulating chondrocytes proliferation Y. Chen, J. Ke, X. Long,

MicroRNA-92a-3p regulates the expression of cartilage-specific genes by directly targeting histone deacetylase 2 in chondrogenesis and degradation G.

MicroRNA-92a-3p regulates the expression of cartilage-specific genes by directly targeting histone deacetylase 2 in chondrogenesis and degradation G.

Histone deacetylase inhibitors suppress mechanical stress-induced expression of RUNX-2 and ADAMTS-5 through the inhibition of the MAPK signaling pathway.

M. M. J. Caron, P. J. Emans, M. M. E. Coolsen, L. Voss, D. A. M

Impaired glycolytic metabolism causes chondrocyte hypertrophy-like changes via promotion of phospho-Smad1/5/8 translocation into nucleus T. Nishida,

Synovial mesenchymal stem cells from osteo- or rheumatoid arthritis joints exhibit good potential for cartilage repair using a scaffold-free tissue engineering.

TGFβ inhibition during expansion phase increases the chondrogenic re-differentiation capacity of human articular chondrocytes R. Narcisi, L. Signorile,

Hypertrophic differentiation during chondrogenic differentiation of progenitor cells is stimulated by BMP-2 but suppressed by BMP-7 M.M.J. Caron, P.J.

X. Zhang, I. Prasadam, W. Fang, R. Crawford, Y. Xiao

Cannabinoid WIN-55,212-2 mesylate inhibits interleukin-1β induced matrix metalloproteinase and tissue inhibitor of matrix metalloproteinase expression.

Subtractive gene expression profiling of articular cartilage and mesenchymal stem cells: serpins as cartilage-relevant differentiation markers S. Boeuf,

H. Bretschneider, M. Stiehler, A. Hartmann, E. Boger, C. Osswald, J

J.E. Lafont, F.-A. Poujade, M. Pasdeloup, P. Neyret, F. Mallein-Gerin

Restriction of spontaneous and prednisolone-induced leptin production to dedifferentiated state in human hip OA chondrocytes: role of Smad1 and β-catenin.

Sequential exposure to fibroblast growth factors (FGF) 2, 9 and 18 enhances hMSC chondrogenic differentiation D. Correa, R.A. Somoza, P. Lin, S. Greenberg,

Effect of glucosamine and its peptidyl-derivative on the production of extracellular matrix components by human primary chondrocytes D. Stoppoloni, L.

Endoplasmic reticulum stress-induced apoptosis contributes to articular cartilage degeneration via C/EBP homologous protein Y. Uehara, J. Hirose, S.

K.A. Payne, D.M. Didiano, C.R. Chu Osteoarthritis and Cartilage

M. Akagi, M. D. , Ph. D. , A. Ueda, M. D. , T. Teramura, Ph. D. , S

G.-I. Im, H.-J. Kim Osteoarthritis and Cartilage

MicroRNA-320 regulates matrix metalloproteinase-13 expression in chondrogenesis and interleukin-1β-induced chondrocyte responses F. Meng, Z. Zhang, W.

Consequences of chondrocyte hypertrophy on osteoarthritic cartilage: potential effect on angiogenesis L. Pesesse, C. Sanchez, J.-P. Delcour, A. Bellahcène,

CaMKII inhibition in human primary and pluripotent stem cell-derived chondrocytes modulates effects of TGFβ and BMP through SMAD signaling B. Saitta,

Comparative analysis with collagen type II distinguishes cartilage oligomeric matrix protein as a primary TGFβ-responsive gene H. Li, D.R. Haudenschild,

Glucosamine promotes chondrogenic phenotype in both chondrocytes and mesenchymal stem cells and inhibits MMP-13 expression and matrix degradation A.

CRISPR-Cas9 targeting of MMP13 in human chondrocytes leads to significantly reduced levels of the metalloproteinase and enhanced type II collagen accumulation

Differentiation potential of human muscle-derived cells towards chondrogenic phenotype in alginate beads culture R. Andriamanalijaona, Ph.D., E. Duval,

H.H. Lee, M.J. O'Malley, N.A. Friel, C.R. Chu

M. E. R. van Meegeren, G. Roosendaal, N. W. D. Jansen, M. J. G

M. Stumm, E. Boger, C. G. Gaissmaier, C. Oßwald, M. Blankenburg, R. D

V. Queirolo, D. Galli, E. Masselli, R. M. Borzì, S. Martini, F

Osteoblasts from the sclerotic subchondral bone downregulate aggrecan but upregulate metalloproteinases expression by chondrocytes. This effect is mimicked.

Enrichment of committed human nucleus pulposus cells expressing chondroitin sulfate proteoglycans under alginate encapsulation Y. Sun, M. Lv, L. Zhou,

M.M.-G. Sun, F. Beier Osteoarthritis and Cartilage

Expression of the semicarbazide-sensitive amine oxidase in articular cartilage: its role in terminal differentiation of chondrocytes in rat and human

E. Farrell, P. Wielopolski, P. Pavljasevic, N. Kops, H. Weinans, M. R

Single cell sorting identifies progenitor cell population from full thickness bovine articular cartilage Y. Yu, H. Zheng, J.A. Buckwalter, J.A. Martin

H Jo, M. D. , J. S Park, Ph. D. , E. M Kim, M. S. , M. Y Jung, Ph. D

Synovial mesenchymal stem cells from osteo- or rheumatoid arthritis joints exhibit good potential for cartilage repair using a scaffold-free tissue engineering.

Enhancing and maintaining chondrogenesis of synovial fibroblasts by cartilage extracellular matrix protein matrilins M. Pei, M.D., Ph.D., J. Luo, M.D.,

Synergistic effects of growth and differentiation factor-5 (GDF-5) and insulin on expanded chondrocytes in a 3-D environment B. Appel, J. Baumer, D.

Autologous chondrocyte implantation (ACI) for aged patients: development of the proper cell expansion conditions for possible therapeutic applications

J. Ranstam Osteoarthritis and Cartilage

Overexpression of hsa-miR-148a promotes cartilage production and inhibits cartilage degradation by osteoarthritic chondrocytes L.A. Vonk, A.H.M. Kragten,

Synergistic effect of chondroitin sulfate and cyclic pressure on biochemical and morphological properties of chondrocytes from articular cartilage G.

G. van den Akker, T. Welting, D. Surtel, A. Cremers, W. Voncken, L

Regulation of senescence associated signaling mechanisms in chondrocytes for cartilage tissue regeneration S. Ashraf, B.-H. Cha, J.-S. Kim, J. Ahn, I.

Chondroitin sulfate modulation of matrix and inflammatory gene expression in IL-1β- stimulated chondrocytes – study in hypoxic alginate bead cultures

Identification of molecular markers for articular cartilage

Viperin; A novel chondrogenic regulator

Battling low back pain with novel NP cell models

Proteome characterization of human articular chondrocytes leads to novel insights in the function of small heat-shock proteins in chondrocyte homeostasis

R. H. J. Das, M. Sc. , H. Jahr, Ph. D. , J. A. N. Verhaar, M. D. , Ph

Low oxygen tension stimulates the redifferentiation of dedifferentiated adult human nasal chondrocytes1 1 Supported by IsoTis S.A. J. Malda, Ph.D., C.A.

Optimal ratio of adipose stem cells and bone marrow stem cell to promote osteogenic differentiation and angiogenesis G.-I. Im, K.-I. Kim, N.-H. Jo Osteoarthritis.

R.D. Chavez, G. Coricor, J. Perez, H.-S. Seo, R. Serra

Peroxisome proliferator activated receptor alpha activation decreases inflammatory and destructive responses in osteoarthritic cartilage S. Clockaerts,

Chondro-protective effects of low intensity pulsed ultrasound

Comparative study of depth-dependent characteristics of equine and human osteochondral tissue from the medial and lateral femoral condyles J. Malda,

Structured three-dimensional co-culture of mesenchymal stem cells with chondrocytes promotes chondrogenic differentiation without hypertrophy M.E. Cooke,

Effect of expansion medium on ex vivo gene transfer and chondrogenesis in type II collagen–glycosaminoglycan scaffolds in vitro R.M. Capito, Ph.D., M.

Presentation transcript:

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 1170-1178 (October 2012) DOI: 10.1016/j.joca.2012.06.016 Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

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 2012 20, 1170-1178DOI: (10.1016/j.joca.2012.06.016) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

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 2012 20, 1170-1178DOI: (10.1016/j.joca.2012.06.016) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions

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 2012 20, 1170-1178DOI: (10.1016/j.joca.2012.06.016) Copyright © 2012 Osteoarthritis Research Society International Terms and Conditions