Isolation and Characterization of Mesenchymal Stem Cells Derived from Human Amniotic Membrane by Explant Technique

Background: Mesenchymal stem cells (MSCs) are adult stem cell types. When examined by phase contrast microscopy, they appear spindle-shaped and fibroblast-like cell aggregates. Up to now, MSCs have been isolated from many tissues such as bone marrow, adipose tissue, cartilage, placenta, amniotic membrane and umbilical cord. Our aim in this study was to isolate mesenchymal stem cells from amniotic membrane by explant culture technique easily and without needing a large amount of equipment. Materials and Methods: In the cell culture laboratory, amniotic membranes were dissected into small pieces and explant culture medium was created. Then, flow cytometry analysis was performed at passage 3 (P3) to determine the characterization of proliferating cells. Differentiation experiments were conducted to show the changes in adipogenic, chondogenic and osteogenic direction. Results: In our study, mesenchymal stem cells were isolated from the human amniotic membrane by explant cell culture technique. Flow cytometry analysis showed that the cells expressed mesenchymal stem cell markers (CD73 and CD90) but not hematopoietic stem cell markers (CD34). In differentiation experiments, adipogenic, chondrogenic and osteogenic changes were observed. Conclusions: After the widespread use of mesenchymal stem cells isolated from the umbilical cord as a valuable source of regenerative and reparative medicine, studies on the isolation of mesenchymal stem cells from the amniotic membrane have intensified.