SHEEP AMNIOTIC FLUID CELLS (SAFCS) DIFFERENTIATION TOWARDS AN OSTEOGENIC PATHWAY

Objectives: The aim of the our study was to evaluate how sheep amniotic fluid could be a potential source for stem cells able to differentiate into osteogenic pathways. Methods: Amniotic fluid samples (about 5ml) were withdrawn from sheep in pregnancy. Each sample contains a cell number ranging from 2x103 to 2x106. The pellets of amniotic fluid were directly resuspended in osteogenic medium without the previous selection of Amniotic Fluid Mesenchymal Stem Cells. At established times, the expression of typical osteogenic markers was investigated by RT-PCR. Moreover, presence of calcium deposition in cell culture was observed at light microscope after Alizarin Red staining. Results: sAFCs isolated and treated according to this protocol are able to produce nodules of calcium mineralization within 18 days from withdrawal. After 30 days from withdrawal, the cluster of cells showed at RT-PCR analyses a full expression of osteogenic markers (COL1, ONC, OPN, OCN, OPG, BSP, Runx2). Moreover at the same time, the sample was investigated at light microscope to highlight the presence of calcium deposition with Alizarin Red staining. Conclusion: Our research showed osteoblast-like cells differentiation obtained from sheep amniotic fluid. The presence of calcium phosphate granules around osteoblast-like cells suggests the potential role of these stem cells population to grow and repair pathological or genetic deseases. Further investigation may be necessary to better understand the mechanisms outstanding tissues engineering and bone repair treatment.