Issue title: Selected papers of the 5th International Symposium on Mechanobiology of Cartilage and Chondrocyte, Athens, May 2007
Article type: Research Article
Authors: Cournil-Henrionnet, Christel; | Huselstein, Céline; | Wang, Yun | Galois, Laurent; | Mainard, Didier; | Decot, Véronique | Netter, Patrick | Stoltz, Jean-François; | Muller, Sylvaine | Gillet, Pierre; | Watrin-Pinzano, Astrid
Affiliations: UMR 7561 CNRS, Université Nancy I, F 54505 Vandoeuvre-les-Nancy, France | UMR 7563 CNRS, UHP-INPL, F 54505 Vandoeuvre-les-Nancy, France | Chirurgie Orthopédique et Traumatologique, F 54035, Chu Nancy, France | Unité de Thérapie Cellulaire et Tissulaire, F 54505 Chu Nancy-Brabois, France
Note: [] Equal contribution to this work.
Note: [] Equal contribution to this work.
Note: [] Address for correspondence: Prof. Pierre Gillet, UMR 7561 CNRS, Nancy Université, Physiopathologie et Pharmacologie Articulaires, Faculté de Médecine, BP 184, Avenue de la Forêt de Haye, F 54505 Vandoeuvre-les-Nancy, France. E-mail: p.gillet@chu-nancy.fr.
Abstract: Both chondrocytes and mensenchymal stem cells (MSCs) are the most used cell sources for cartilage tissue engineering. However, monolayer expansion to obtain sufficient cells leads to a rapid chondrocyte dedifferentiation and a subsequent ancillary reduced ability of MSCs to differentiate into chondrocytes, thus limiting their application in cartilage repair. The aim of this study was to investigate the influence of the monolayer expansion on the immunophenotype and the gene expression profile of both cell types, and to find the appropriate compromise between monolayer expansion and the remaining chondrogenic characteristics. To this end, human chondrocytes, isolated enzymatically from femoral head slice, and human MSCs, derived from bone marrow, were maintained in monolayer culture up to passage 5. The respective expressions of cell surface markers (CD34, CD45, CD73, CD90, CD105, CD166) and several chondrogenic-related genes for each passage (P0–P5) of those cells were then analyzed using flow cytometry and quantitative real-time PCR, respectively. Flow cytometry analyses showed that, during the monolayer expansion, some qualitative and quantitative regulations occur for the expression of cell surface markers. A rapid increase in mRNA expression of type 1 collagen occurs whereas a significant decrease of type 2 collagen and Sox 9 was observed in chondrocytes through the successive passages. On the other hand, the expansion did not induced obvious change in MSCs gene expression. In conclusion, our results suggest that passage 1 might be the up-limit for chondrocytes in order to achieve their subsequent redifferentiation in 3D scaffold. Nevertheless, MSCs could be expanded in monolayer until passage 5 without loosing their undifferentiated phenotypes.
Keywords: Human mesenchymal stem cells, bone marrow, chondrocytes, cartilage engineering, gene expression, flow cytometry
DOI: 10.3233/BIR-2008-0487
Journal: Biorheology, vol. 45, no. 3-4, pp. 513-526, 2008
