The aim of this study was to isolate and characterize rat Adipose Derived Mesenchymal Stem Cells (AD-MSCs) in order to
evaluate the stemness markers gene expression levels, together with the assessment of phenotypic expression of the CD90
and CD45 markers. MSCs were obtained from subcutaneous adipose tissue of 16 Wistar rats. Microbiological controls were
performed to exclude the presence of bacteria, fungi and viruses in tissue. Adipose tissue was mechanically and enzimatically
fragmented and stromal cell fraction was seeded in adherent culture flasks in DMEM 20% FBS. After 48h, the medium
was replaced. Cells were characterized by evaluating their ability to adhere to the plastic; the clonogenic potential by Colony
Forming Unit (CFU) assay and their ability to differentiate adipocytes and osteocytes. AD-MSCs were analysed for the
expression of three stem cell master genes (OCT-4, SOX-2 and NANOG) at different cell subcultures by Sybr Green RealTime
PCR. Statistical analysis, performed with REST software, showed that the expression of the genes is maintained from
subculture 0 to 5. Moreover, flow cytometry analysis confirmed the mesenchymal nature of cells isolated from adipose tissue
as they were positive for CD90, which is a MSCs surface marker, and negative for CD45 (typical hematopoietic marker)
