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3T3-L1 differentiation is an economical and convenient way to generate adipocyte-like cells for experiments.
Preparation of media must be carried out in a tissue culture hood under aseptic conditions. MDI (methylisobutylxanthine, dexamethasone, insulin) induction medium and insulin medium should be freshly prepared.
Preparation of MDI induction medium
Preparation of insulin medium
Differentiation into adipocyte-like cells can be tracked by Oil Red O staining to monitor lipid accumulation, or by monitoring the expression of adipocyte markers such as adiponectin and FABP4.
Protocol adapted from Reed BC, Lane D (1980). Insulin receptor synthesis and turnover in differentiating 3T3-L1 preadipocytes. Proc. Natl. Acad. Sci. USA. 77, 285–289