INTRODUCTION: ADSCs are at the forefront of stem cell therapy due to less invasive isolation procedures and its higher yield in cell number after purification. Growth Factors (GFs) such as Transforming Growth Factor β (TGF-β) and Bone Morphogenetic Proteins (BMPs) have been investigated as effective tools for driving tenogenic differentiation of stem cells. For example, recent studies highlighted the promising effect of BMP-12 in the tenogenic differentiation of ADSCs . Other studies have compared the effect of different GFs in the upregulation of key markers involved in tendon commitment both in ADSCs  or other stem cell lines . Herein, we focused on the characterisation and the differentiation potential of human ADSCs in the presence of several GFs in the absence of serum. METHODS: ADSCs were serum starved and treated with different customised media as the differentiation factor. After specific time points, several techniques were performed: 1) Analysis of messenger RNA (mRNA) for common tendon markers was performed by Real-Time Polymerase Chain Reaction (RT-PCR). Likewise, markers for other cell lineages were analysed. 2) Protein expression was confirmed by Western blotting. 3) Cellular localisation of the markers was evaluated by Immunocytochemistry (ICC) techniques. RESULTS & DISCUSSION: In this study, we evaluated the tenogenic effect of different GFs on ADSCs. Scleraxis, a transcription factor involved in tendon development, was upregulated at the mRNA level, but the expression was significantly different depending on the GFs used. Collagen type 1, the most abundant type in tendon, was greatly induced. Interestingly, deposition of Collagen I on the extracellular matrix (ECM) by the ADSCs was altered depending on the media used, which was confirmed by ICC. Additionally, cell morphology and confluency changed depending on the GF used. CONCLUSIONS: Our data suggest that specific environments may be needed for the tenogenic development and potential application for the treatment of the injuries without overstimulation. Overall, we believe ADSCs represent a promising approach for tendon regeneration and repair. ACKNOWLEDGEMENTS: Financial support was received from Rosetrees Trust Fund, Arthritis Action UK and University of East Anglia, Faculty of Science.
|Published - 27 Jun 2019