The differentiated mouse 3T3-L1 adipocytes (3T3-L1 model) were used in studying glucose metabolisms without the need for feeding (Sprague-Dawley, SD model) the rat prior to hypoglycemic activity evaluation. Both models were adopted to evaluate the hypoglycemic activities of 58 yeast strains isolated from various sources (grape, vine yard soil, winery soil). Among the 58 tested yeast isolates, strain 54 (Saccharomyces pastorianus no. 54) which showed the highest hypoglycemic activity was chosen to be the test strain. The optimal insulin concentration used in these 2 models (SD and 3T3-L1) for measuring the hypoglycemic activity of hypoglycemic yeast extract (HGYE) was 10 nM. The range of linear relation in the dose-response curve was 0-1000 g/ml for SD model, and 0-250 g/ml for 3T3-L1 model. The linear coefficient was 0.8611. The radioactive labeled 2-[1-14C]-Deoxy-D-Glucose was also used to confirm cytoplasmic glucose uptake by 3T3-L1 adipocytes. Comparing both the results of insulin effect and dose response of HGYE by both models, it was concluded that the 3T3-L1 model can serve as a rapid and reliable assay model for in vitro evaluation of hypoglycemic activity of yeast.Key words: 3T3-L1 adipocytes, Sprague-Dawley (SD) rat, epididymal adipocytes, hypoglycemic activity, yeast