Isolation and Culture of Mesenchymal Stem Cells From Rabbit Scapular Subcutaneous Adipose Tissue and Their Ability to Differentiate Into Osteoblasts

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. 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. 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. 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. 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 differentiation ability of MSCs in normal medium and hope to employ these cells for the regeneration of damaged bone tissues in the future.