Stromal-vascular(S-V) cells isolated from omental, subcutaneous, intermuscular, and intramuscular adipose tissues of Korean native steers by collagenase digestion were used to evaluate the effects of various hormones on the differentiation in culture. The cells were seeded at a density of 3 × 10⁴ cells/㎠ on the tissue culture plates in M199 medium plus 20% fetal calf serum(FCS) and cultured at 37℃ under a humidified atmosphere of 95% air : 5% CO₂ for about 7 days. After reaching confluence, the cells were treated with DMEM/F12 medium containing 10% FCS and either insulin, triiodothyronine or dexamethasone for induction of adipose differentiation for 2∼4 weeks, and thereafter cellular glycerol-3-phosphate dehydrogenase(GPDH) acitivity was determined as an index of adipocyte differentiation. GPDH activity was about 5 times higher in intermuscular adipose cell cultured in DMEM/F12 medium containing insulin and lipid than in medium containing only insulin, suggesting that lipid plays an important role in development of internmscular triglyceride accumulation. Differentiation in the intermuscular adipose cell was induced by insulin, but the subcutaneous and intramuscular adipose cells did not respond to insulin. Very a few GPDH activity was observed in the omental adipose cell in comparision to the intermuscular adipose cell. Neither triiodothyronine nor dexamethasone were related to adipogenic activity. More GPDH activity was measured in intermuscular adipose cell cultured in insulin plus triiodothyronine(T₃) than in insulin alone, suggesting that T₃ is related to enhance the level of GPDH activity during differentiation and treatment with insulin plus T₃ are responsible for successful adipocyte differentiation in intermuscular adipose cell. The differences in hormone responses of cells derived from different locations of adipose tissue indicated that differences may exist in the regulation of the differentiation of preadipocyte from different locations.