Affiliations: INSERM U791, LIOAD, Group “STEP”, Nantes University, Nantes, France | National Veterinary School, CRIP – Experimental Surgery Department, Nantes, France | Graftys SA, Aix en Provence, France | INRA UMR703, National Veterinary School, Nantes, France
Note: [] Address for correspondence: Jérôme Guicheux, INSERM U791, Laboratoire d'Ingénierie Ostéo-Articulaire et Dentaire (LIOAD), Faculté de Chirurgie Dentaire, 1 Place Alexis Ricordeau, 44042 Nantes Cedex 1, France. Tel.: +33 240412919; Fax: +33 240083712; E-mail: jerome.guicheux@inserm.fr.
Abstract: Articular cartilage does not repair itself spontaneously. To promote its repair, the transfer of stem cells from adipose tissue (ATSC) using an injectable self-setting cellulosic-hydrogel (Si-HPMC) appears promising. In this context, the objective of this work was to investigate the influence of in vitro chondrogenic differentiation of ATSC on the in vivo cartilage formation when combined with Si-HPMC. In a first set of experiments, we characterized ATSC for their ability to proliferate, self renew and express typical mesenchymal stem cell surface markers. Then, the potential of ATSC to differentiate towards the chondrogenic lineage and the optimal culture conditions to drive this differentiation were evaluated. Real-time RT-PCR and histological analysis for sulphated glycosaminoglycans and type II collagen revealed that 3-dimensional culture and hypoxic condition favored ATSC chondrogenesis regarding mRNA expression level and the corresponding proteins production. In order to assess the phenotypic stability of chondrogenically-differentiated ATSC, real-time RT-PCR for specific terminal chondrogenic markers and alkaline phosphatase activity assay were performed. In addition to promote chondrogenesis, our culture conditions seem to prevent the terminal differentiation of ATSC. Histological examination of ATSC/Si-HPMC implants suggested that the in vitro chondrogenic pre-commitment of ATSC in monolayer is sufficient to obtain cartilaginous tissue in vivo.
