Isolation, characterization, and in vitro differentiation of osteogenic human amniotic fluid- derived stem cells

Stem cells (SCs) regenerative therapy represents an emerging strategy for the treatment of human diseases. Mesenchymal stem cells (MSCs) offer significant promise as a multipotent source for cell-based therapies and could form the basis for the differentiation and cultivation of tissue grafts to replace damaged tissue. The main goal of this study was to focus on MSCs and to analyze their differentiation capacity into osteogenic cells. To achieve this aim, MSCs were isolated from amniotic fluid-derived cells (AFSCs). The expression of CD29, CD90, CD105, CD13, CD34, CD14, and Oct-4 of MSCs were characterized by flow cytometry. The proliferation and differentiation of MSCs into osteogenic were examined in vitro. Osteogenic differentiation was determined by staining with Alizarin Red S and real time polymerase chain reaction (RT-PCR) analysis of genes markers expression after 4, 7, 14, and 21 days. Our results revealed that the patterns of CD expression markers were highly positive for CD29 (50%), CD 90 (79.1%), CD 105 (79.5%), CD13 (36.5%) and Oct-4 (31.4%) and negative for CD14 (11.3%) and CD34 (15%). MSCs showed proliferative potential and were capable of osteogenic differentiation characterized by Alizarin Red S staining and expression of molecular detection of genes. MSCs from amniotic fluid had the ability to differentiate in vitro into osteogenic under specific culture conditions. AFSCs may represent sources of characterized pluripotent SCs that can be easily collected and amplified in vitro. These MSCs may be used in preclinical studies on large animals to develop future human therapies.