Isolation and Morphological Characterization of Ovine Adipose-Derived Mesenchymal Stem Cells in Scanning Electron Microscopy (SEM)

The main purpose of this study is to provide advanced insights into scanning electron microscopy (SEM) of adipose derived mesenchymal stem cells (oAD-MSCs). In this study after isolation and proliferation of ADMSCs, their cell surface markers characterized using antibodies RT-PCR. SEM was used to study of ultrastructure of oAD-MSCs. Adipose tissue was obtained from tail fat of sheep. Surface markers (CD44, CD90, CD34, CD31) evaluated by RT-PCR. RT-PCR was done for CD44, CD90, CD34 and CD31 to identify of cells. Morphological characterization was done by inverted microscope and SEM. Cells was prepared by glutaraldehyde for first fixation and tetroxide osmium for second fixation and dehydration with different percent of ethanol. Finally, cells coated with gold and observed in SEM. Adipose derived mesenchymal stem cells (AD-MSCs) were isolated and proliferated. They were positive for CD44 and CD90 markers and negative for CD31 and Cd34 markers in RT-PCR technique. AD-MSCs showed a fibroblast-like, spindleshaped morphology after they attached to the culture flasks observing in inverted microscope. Explanted specimens were imaged with scanning electron microscopy (SEM). SEM provides the main technology to visualize surface features. In SEM the outer surface of the mesenchymal cells could be observed; so, in this study, the pseudopods arising from each cell and extending through each other were clearly shown.