Abstract: Objective To investigate the optimal conditions for establishing insulin-resistant 3T3-L1 adipocytes. Methods
Dexamethason (DEX), 3-isobutyl-methylxanthine (IBMX) and different concentrations of insulin (10-8, 10-7, and 10-6 mol ·L-1)
were used to induce 3T3-L1 preadipocytes into mature adipocytes identified by oil red O staining. We established insulinresistant
3T3-L1 adipocytes cell model (IR-3T3-L1) by exposing the cells to 1 μmol·L-1 DEX, and the changes of glucose concentration
in the cell culture were determined by glucose oxidase-peroxidase (GOD-POD) assay. Results Treatment of 3T3-L1 cells
with DEX, IBMX and 10-6 mol·L-1 insulin for 9 days resulted in the differentiation of >90% of the cells into mature adipocytes.
IR-3T3-L1 cells cultured for 96 h in the culture media containing 1 μmol ·L-1 DEX showed significantly increased glucose
consumption (P=0.0003) as compared with the control group at 36 h (P<0.001). Conclusion 3T3-L1 cells can be induced into
mature adipocytes by exposure to 1 μmol·L-1 DEX, 0.5 mmol·L-1 IBMX and 10-6 mol·L-1 insulin. A 96 h exposure to 1 μmol·L-1
DEX can induce 3T3-L1 adipocytes to acquire insulin resistance that can be maintained for 36 h.