ECTS2013 Poster Presentations Cell biology: osteoblasts and bone formation (50 abstracts)
Evaluation of bone formation capacities of human adipose-derived stromal cells cultured in platelet growth factor-enriched plasma medium.
Fabien Guilloton 1 , Vahideh Rabani 1 , Meadhbh Brennan 2 , Giulio Bassi 3 , Mauro Krampera 3 , Pierre Layrolle 2 , Luc Sensebé 1 & Frédéric Deschaseaux 1
1StromaLab, UMR CNRS 5273, EFS PM, UPS, INSERM U1031, Toulouse, France; 2INSERM U957-LPRO, Nantes, France; 3Stem Cell Research Laboratory, University of Verona, Verona, Italy.
Human adipose-derived stromal cells (ASCs) exhibit strong plasticity and proliferation potentials. In addition, ASCs are easy to harvest and are found at high frequency in adipose tissue samples. This gives us opportunities for their use in bone regeneration therapy. We thus evaluated the bone formation potential of ASCs in vitro and in vivo.
ASCs were isolated from subcutaneous adipose tissue (following Local Ethical Guideline and after patient informed consent) and expanded in vitro in medium containing either 10% fetal calf serum (FBS) or 2% platelet growth factor-enriched plasma (PGP). Cells were then subjected for osteoblastic differentiation by using osteogenic medium containing bone morphogenetic protein 4 (BMP4), β-glycerophosphate, and ascorbic acid. Bone marrow mesenchymal stromal cells (MSCs) were also used for comparison. We chose RUNX2, DLX5, and OSX/SP7 as transcription factors and PTHR1, ALPL, BGLAP as well as calcium deposition capacities as functional read-out to assess in vitro osteoblastic differentiation.
Before differentiation basal expressions of RUNX2 and ALPL proteins were strongly increased in PGP-derived ASCs when compared to 10% FBS–ASCs. After induction of differentiation, phenotypic and functional analyses showed that 2% PGP-derived ASCs were more prone to differentiate into osteoblastic cells than 10% FBS–ASCs. Besides these in vitro studies, bTCP discs were loaded by PGP–ASCs and MSCs alone or in combination (90% MSCs/10% ASCs and 10% MSCs/90% ASCs) and then inserted in the back of Nude mice. After 8 weeks, transplants were harvested and analysed for evaluation of bone formation. Only transplant containing 100 or 90% MSCs contained new bone. On the contrary, ASCs formed fibrous tissue. Therefore, ASCs were able to differentiate into osteoblastic cells in vitro but were not spontaneously capable to induce bone formation in vivo. Better pre-conditioning protocols should solve such defect.
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Bone Abstracts
ISSN 2052-1219 (online)
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