DKK1AND FRZB are necessary for chondrocyte (RE) differentiation and prevention of cell hypertrophy in 3D cultures of human chondrocytes and human mesenchymal stem cells

L Zhong, X Huang, E Rodrigues, J Leijten, T. Verrips, M. El Khattabi, M Karperien, J Post

Research output: Contribution to journalMeeting AbstractOther research output

64 Downloads (Pure)

Abstract

and differentiation was induced by keeping the cells for 21 days in a-MEM supplemented with 10% FBS, 1% sodium pyruvate, 1% antibiotic-antimycotic, 10mM b-glycerophosphate (BGP) and 50mg/ml ascorbic acid-2-phosphate (AA). Alkaline phosphatase activity (ALP) was meas-ured and mineralization was analyzed by alizarin red staining. Gene expression of extracellular matrix genes, genes linked to mineralization and TGFb superfamily genes was analyzed by RT-qPCR. Intracellular signaling pathways (PKCa, ERK1/2, SMAD1/5/8, p38MAPK) were investigated by Western blot. HPDCS were cultured for 21 days in DMEM (þ the same supplements as for MC3T3 differentiation and 100nM dexamethasone). HUVECS were stimulated in EGM-2 medium with 200 ng/ml of BMP6, to trigger endothelial to mesenchymal tran-sition. Then cells were cultured as hPDCS. hPDCs and HUVECS were cultured with supernatants harvested from control, Smoc2þ and DCaBD MC3T3 cells. Results: Mineralization and ALP activity was reduced in Smoc2þ MC3T3 cells compared to controls. Gene expression analysis showed an overall altered differentiation when overexpressing Smoc2. The acti-vation status of PKCa, ERK1/2, p38MAPK and SMAD1/5/8 was sig-nificantly modified in Smoc2þ cells. On the other hand, we could not observe an effect on osteogenesis when silencing Smoc2. DCaBD cells
Original languageEnglish
Article number229
Pages (from-to)S142
JournalOsteoarthritis and cartilage
Volume24
Issue numbersupplement 1
DOIs
Publication statusPublished - Apr 2016
Event2016 OARSI World Congress on Osteoarthrtis - Amsterdam RAI, Amsterdam, Netherlands
Duration: 31 Mar 20163 Apr 2016

Fingerprint Dive into the research topics of 'DKK1AND FRZB are necessary for chondrocyte (RE) differentiation and prevention of cell hypertrophy in 3D cultures of human chondrocytes and human mesenchymal stem cells'. Together they form a unique fingerprint.

Cite this