Background and Objectives:Amniotic membrane (AM) is a very attractive source of Mesenchymal Stem Cells (MSCs) due to the fact that this fetal tissue is usually discarded without ethical conflicts, leading to high efficiency in MSC recovery with no intrusive procedures. The aim of the present work is to establish the method of isolation and purification of human amniotic mesenchymal stem cells (hAMSCs) with characterization of such cells. The placentas included in the study were derived from pregnancies with either a normal evolution or after a caesarian section in full term delivery. The isolation and the culture of cells from the amniotic membrane were followed by the determination of the markers of these cells and evaluation of their gene expression to detect the pluripotential properties of these cells. Design and Methods:MSCs were isolated from human amniotic membrane by collagenase-accutase digestion, and cultured in Dulbecco’s modified Eagle’s medium/F12 medium supplemented with 10% fetal bovine serum. Phenotypic characteristics of these cells were analyzed by means of flow cytometry. The gene expression levels were determined by RT- Polymerase Chain Reactions.Results:The cells successfully isolated from human amniotic membrane expressed representative mesenchymal cell surface markers and the assessment of the gene expression evidenced the pluripotential properties of these cells.Interpretation and Conclusions:Here we confirmed that AM, is an abundant source of human MSCs. This study establishes a potential method for isolation of MSCs from human amnion, in vitro culture, and identification. The isolated cells show phenotypic and genotypic characteristics of mesenchymal stem cells.