A pilot study of human mesenchymal stem cells from visceral and sub-cutaneous fat tissue and their differentiation to osteogenic phenotype.

Abstract

To evaluate the different behavior of two different human adult adipocytes derived stem cells (hASCs) during proliferation and osteogenic differentiation. Human adult adipocytes stem cells (hAT-SCs) from visceral (hAV-SCs) and subcutaneous (hAS-SCs) sites were obtained after surgery procedures of seven patients. All samples were fully investigated and the different proliferation rates were evaluated. All MSCs clusters were cultured with an osteogenic and adipogenic differentiation medium. Homogeneous pools of Mesenchymal Stem Cells (MSCs) were confirmed by Flow-Cytometry Analysis (FACS) and Spectrophotometric Assay. The differentiated cells were eventually assessed for the expression of Alkaline Phosphatase (ALP), Alizarin Red (AR) and Oil Red-O (OR-O) detection, and analyzed by the Spectrophotometric Assay. After osteogenic differentiation, the cell clusters were incubated and analyzed with Real Time-Polymerase Chain Reaction (qRT-PCR) and fluorescence microscopy. The FACS analysis performed on hAT-SCs confirmed the homogenous presence of MSCs in all samples. The ALP, AR stain confirmed the osteogenic differentiation capacity of MSCs towards osteoblast-like-cells. The colorimetric cell metabolic activity (MTS) assay showed an increase in the proliferation rate with different values in both sets hAS-SCs vs. hAV-SCs. These in vitro findings of both hAS-SCs and hAV-SCs suggested an important role of these stem cells for future clinical use in bone regeneration. Indeed, the final outcomes suggested a better performance of cells coming from subcutaneous adipose tissue vs. those from visceral fat tissue.