ADIPOCYTE STEM CELLS BEHAVIOUR DURING OSTEOGENIC DIFFERENTIATION

The aim of the study was to evaluate the different behavior of Adipocyte Stem Cells from sub-cutaneous and visceral sites during proliferation and osteogenic differentiation.

Method:

Human Adipocyte stem cells were taken in 10 pz after surgerical procedures. Samples were selected in homogeneous mesenchymal pool with FACS analysis and SEM and optical microscope were analyzed before and after confluence.The samples were cultured in osteogenic medium and the differentiation was evaluated with Alizarin Red detection. Before and after differentiation, cell clusters were evaluated with RT-PCR, proliferation test and Western Blot analysis.

Result:

All samples from human adipocyte showed a homogenous population of mesenchymal stem cells. The osteogenic differentiation were confirmed with positive Alizarine Red staining analysis. RT-PCR analysis in undifferentiated/differentiated cell clusters were observed the different behaviour of ALP marker. The MTS Assay data showed an increase to 8 days of 70% of the proliferation speed in the differentiated cells. The Western Blot analysis showed an increase of ALP and the absence of PECAM-1markers. FACS analysis at the end of stem cells differentiation in osteoblasts like cells showed a decrease of the CD90 proteins, not associated with variations of the other investigated mesenchymal markers.

Conclusion: The ALP increase and the absence of PECAM-1 protein confirmed the differentiation in osteogenic cluster and not in hematopoietic sense and suggests that the proliferation mechanism not reduced the differentiation processes. The positive Alizarine red analysis and the negative results of red oil staining during human osteoblasts like cells differentiation confirmed the potential role of adypocite and dental pulp stem cells in human tissue engineering.