Abstract:

BACKGROUND: Adipose-derived stem cells (ADSCs) and demineralized bone matrix (DBM) can be obtained from the same donator. However, there are few studies on the co-culture of ADSCs as seed cells while DBM as scaffold.  
OBJECTIVE: To observe the growth and proliferation of ADSCs cultured combining with DBM.
METHODS: The subcutaneous adipose tissue was cut and collected from the inguinal fat pads of rabbits under aseptic condition. Primary ADSCs were isolated subsequently by the methods of stirring digestion within collagenase type Ⅰ. After primary culture and subculturing, ADSCs were expanded to the 3rd passage. Cell morphology and growth were observed under an inverted microscope, and the expression of cell surface antigen was detected using immunofluorescence staining. DBM was prepared using rabbit ilium. The 3rd passage of ADSCs was seeded into the allogenic DBM scaffold sample for co-culture. Cell adherence and growth were observed under an inverted microscope at 1 week after co-culture, and the haematoxylin-eosin staining was performed at 2 weeks after co-culture. 
RESULTS AND CONCLUSION: ADSCs isolated from rabbit subcutaneous adipose tissues expanded rapidly. After the third passage, most of the passage cells were positive for CD44 but negative for CD34. The prepared DBM was three-dimensional stereochemical structure with more hollows; there were good plasticity and tolerable mechanical strength. Scanning electron microscopy and haematoxylin-eosin staining revealed that ADSCs adhered and grew well within the DBM scaffolds, and could synthesize cartilage extracellular matrix in the scaffold. These results show that the DBM can provide an appropriate microenvironment for the proliferation and expressions of extracellular matrix of ADSCs.