Abstract: Objective To investigate the method of isolation, culture and identification of human orbital adipose-derived stromal cells (hOADSCs) and it's ability of adipogenic differentiation in vitro. Methods The hOADSCs were isolated by tissue explant culture technique, then were primary cultured and subcultured in vitro. The second generation of hOADSCs was detached, and the expressions of cell surface antigen CD29, CD31, CD44 and CD45 were analyzed with flow cytometry to identify the cells. The hOADSCs were induced to be adipogenesis with the adipogenic induction medium in vitro and stained with oil red O. The morphology of the hOADSC was observed by phase contrast microscopy. The plating efficiency and doubling time of the hOADSCs were tested. The hOADSCs proliferation was detected by CCK-8, and the cell growth curve was made. Results The hOADSCs were isolated by tissue explant culture technique and exhibited a fibroblast- or spindle-like morphology. The hOADSCs grew rapidly after subculturing. The expression rate of cell surface antigen CD29, CD31, CD44 and CD45 was 99.0%, 1.5%, 99.1% and 1.2%, respectively. The hOADSCs were differentiated into adipocytes with the adipogenic induction medium in vitro and the intracellular lipid droplets were stained in red by oil red O. The plating efficiency and doubling time of the subcultured hOADSC were 92.6% and 1.98days, respectively. The hOADSCs could be subcultured for more than ten generations in vitro. Conclusion In this experiment, the hOADSCs were isolated, cultured by tissue explant culture technique and could be differentiated into mature adipocytes with the adipogenic induction medium in vitro.

Key words: Orbit, Adipose-derived stromal cell, Culture