Objective Divided orbital fibroblasts from patients with thyroid-associated ophthalmopathy (TAO) into CD90+ and CD90- subsets respect to surface CD90 expression, then determined whether CD90+ and/or CD90- fibroblasts were capable of differentiating into lipofibroblasts. Methods Fibroblasts subset separation into CD90+ and CD90- subsets was accomplished by three to four rounds of magnetic bead selection, then treated with 3-isobutyl-1-lmethylxanthine (IBMX), insulin and dexamethasone which was an known inducer of the lipofibroblastic phenotype, then the cells were observed every day to find Lipid droplets. Oil red O staining were conducted at 5 d, 10 d, 15 d, 20 d and 25 d after inducing. The percent of lipofibroblasts were calculated. Results The ratio in fibroblast derived from extraocular muscles of differentiating into lipofibroblast is less than from connective/adipose tissue (P+ fibroblast is less than CD90- fibroblast (P+ cells derived from extraocular miscles could not be induced to differentiate into lipofibroblast. The ratio in CD90- fibroblast from connective/adipose tissue is highest (P- connective/adipose tissue.