Anika Jonitz-Heincke1, Annett Klinder1, Diana Boy2, Achim Salamon3, Doris Hansmann1, Juliane Pasold1, Andreas Buettner2, Rainer Bader1. 1. 1 Department of Orthopaedics, Biomechanics and Implant Technology Research Laboratory, University Medical Center Rostock, Rostock, Germany. 2. 2 Institute of Forensic Medicine, University Medical Center Rostock, Rostock, Germany. 3. 3 Department of Cell Biology, University Medical Center Rostock, Rostock, Germany.
Abstract
OBJECTIVE: In the present in vitro study, we analyzed the chondrogenic differentiation capacity of human chondrocytes postmortally isolated from unaffected knee cartilage by the addition of transforming growth factor-β1 (TGF-β1) and/or insulin-like growth factor-1 (IGF-1) and different oxygen levels. DESIGN: After 14 and 35 days, DNA concentrations and protein contents of Col1, Col2, aggrecan as well as glycosaminoglycans (GAGs) of chondrocytes cultivated as pellet cultures were analyzed. Additionally, expression rates of mesenchymal stem cell (MSC)-associated differentiation markers were assessed in monolayer cultures. RESULTS: All cultivated chondrocytes were found to be CD29+/CD44+/CD105+/CD166+. Chondrocytic pellets stimulated with TGF-β1 showed enhanced synthesis rates of hyaline cartilage markers and reduced expression of the non-hyaline cartilage marker Col1 under hypoxic culture conditions. CONCLUSIONS: Our results underline the substantial chondrogenic potential of human chondrocytes postmortally isolated from unaffected articular knee cartilage especially in case of TGF-β1 administration.
OBJECTIVE: In the present in vitro study, we analyzed the chondrogenic differentiation capacity of human chondrocytes postmortally isolated from unaffected knee cartilage by the addition of transforming growth factor-β1 (TGF-β1) and/or insulin-like growth factor-1 (IGF-1) and different oxygen levels. DESIGN: After 14 and 35 days, DNA concentrations and protein contents of Col1, Col2, aggrecan as well as glycosaminoglycans (GAGs) of chondrocytes cultivated as pellet cultures were analyzed. Additionally, expression rates of mesenchymal stem cell (MSC)-associated differentiation markers were assessed in monolayer cultures. RESULTS: All cultivated chondrocytes were found to be CD29+/CD44+/CD105+/CD166+. Chondrocytic pellets stimulated with TGF-β1 showed enhanced synthesis rates of hyaline cartilage markers and reduced expression of the non-hyaline cartilage marker Col1 under hypoxic culture conditions. CONCLUSIONS: Our results underline the substantial chondrogenic potential of human chondrocytes postmortally isolated from unaffected articular knee cartilage especially in case of TGF-β1 administration.
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