Abstract
Background: The objectives of this study were to separate and culture mesenchymal stem cells (MSCs) from adipose tissue, examine the
expression of surface markers on these cells, and determine their ability to differentiate into osteoblasts in normal medium.
Objectives: The objectives of this study were to separate and culture mesenchymal stem cells (MSCs) from adipose tissue, examine the
expression of surface markers on these cells, and determine their ability to differentiate into osteoblasts in normal medium.
Materials and Methods: Sterile adipose tissue was obtained from the scapular subcutaneous adipose tissue of two rabbits (average weight,
2.8 kg) for cultivation and differentiation by either liposuction with a blunt hallow tip cannula or by direct surgery. The morphology,
differentiation, and expression of mesenchymal-specific surface markers of rabbit, such as CD90, CD45, CD73, CD44, and CD105, were
examined in cells from the third passage by flow cytometry. The MSCs from adipose tissue were stained with a lentivirus genome for cell
tracking. The differentiation of MSCs into osteoblasts was investigated using a specific histological stain, Alizarin red.
Results: The identity of adipose tissue cells was confirmed by oil-red O staining and examination under an optical microscope at both
the initial stage and after differentiation into mesenchymal cells. The results demonstrated that cells derived from adipose tissue
differentiated into mesenchymal cells. The nature of the mesenchymal cells was confirmed by the expression of specific surface markers,
including CD90, CD45, CD44, CD73, and CD105, by flow cytometry. Finally, Alizarin red staining confirmed the differentiation of MSCs into
osteoblasts.
Conclusions: Based on our findings, we conclude that the separation and reproduction of adipose tissue cells is an appropriate method
for purification of MSCs in animal studies. Regarding the histomorphometric and flow cytometry analysis results, we demonstrated the
different future.