Mouse Mesenchymal Stem Cells Differentiation into the Endothelial Lineage
Abstract
Background. Numerous studies have showed that bone marrow derived mesenchymal stem cells are multipotent and present the capacity to differentiate into various cell types, including osteocytes, adipocytes, chondrocytes and neural cells. Recent studies have reported the stem cells ability to differentiate into vascular endothelial cells in vitro.
Aims. Our objectives were the isolation and the cultivation of bone-marrow- derived mesenchymal stem cells in mice. This was followed by the morphological characterization of primary and secondary culture cells and the highlighting of multipotent properties by endothelial lineage differentiation and immunophenotyping of differentiated cells.
Material and Methods. Mesenchymal stem cells were isolated from CD1 mice and cultured in supplemented IMDM medium. Multipotent potential was evaluated by mesenchymal stem cells differentiation into endothelial-like cells using EMB-2 medium supplemented with EGM2 Sigle Quots. Cells characterization was done by immunophenotyping, using the BD FACS Canto II flow cytometer.
Results. In the 12 th day of culture, the number and density of fibroblast-like cells colonies increased. Cell population shared diverse morphologies: spindle-shaped, star- shaped and hexagonal-shaped cells. Four days after induced differentiation, there were obvious cell islets delimited by empty polygonal areas and interconnected by cellular cords, which is characteristic for differentiated endothelial cells. Immunophenotyping showed a percentage of 67,2% CD105 positive cells and a percentage of 73,5% CD105 and CD31 positive cells. All these suggest an advanced stage of cells differentiation.
Conclusions. To obtain a homogenous cell population of derived endothelial cells from mesenchymal stem cells in mice, we recommend the use of supplemented EMB-2 culture medium.