An Inexpensive and Simple Method for Isolation Mesenchymal Stem Cell of Human Amnion Membrane

Abstract

<strong>Background:</strong> Background: Human-amnion membrane mesenchymal stem cells (hAMSCs) are the accessible cells that can be grown in <em>in vitro</em> condition to produce a great number of cells required for cell therapy in medicine. Several protocols have been proposed for isolation of hAMSCs, but the most of them are multi-step and expensive. The purpose of this article was to introduce a one-step, inexpensive protocol.<br /> <strong>Methods:</strong> In this study human amnion membrane (hAM) was obtained from selected caesarean-sectioned births. The AM was sliced into small pieces and to isolate MSCs, it was digested only with one step instead of multi-step. Isolated cells were cultured in Dulbeco^,s Modified Eagle^,s Medium-Low Glucose (DMEM-LG) with 10% fetal bovine serum (FBS) without adding growth factors. After 80-90% confluency, the adherent cells were characterized by flow cytometry and multi- potentiality differentiation toward adipocyte-, and osteocyte -like cells.<br /> <strong>Results:</strong> The results showed that hAMSCs isolated from hAM expressed CD105, CD90 and CD73 such as other MSCS, but did not express CD34 and CD45 hematopoietic markers. The osteogenic and adipogenic differentiation of the isolated cells were proven by Alizarin Red and Oil-Red-O straining, respectively.<br /> <strong>Conclusions:</strong> The results showed that the stem cells derived from the AM belonged to the mesenchymal stem cells family. Furthermore, this method made it easier and cheaper to obtain this type of stem cells<strong>.</strong>