Monday, October 13, 2008


Fig. 1: 100% confluent of 3T3-L1 preadipocytes



Fig. 2: Mature 3T3-L1 adipocytes


3T3-L1 cell is a cell line derived from disaggregated Swiss 3T3 mouse embryos. The cells are able to undergo adipogenesis to form mature adipocytes. Here, the preadipocytes were cultured with DMEM (Dulbecco’s modified Eagle’s medium) supplemented with 10% fetal calf serum (FCS). Once the cells were 100% confluent (Fig.1), the cells were incubated with additional 48 h before initiating differentiation. At this time, 3T3-L1 preadipocytes expressed very early markers of adipocyte differentiation.

Subsequently, the preadipocytes were induced with a defined adipogenic media to differentiate the cells. The media is DMEM with a combination of dexamethasone, isobutylmethylxanthine, insulin and 10% fetal bovine serum (FBS). 2 days after induction, the isobutylmethylxanthine and dexamethasone were removed but insulin was maintained in media for another 2 days. Thereafter, the media were changed in DMEM with 10% FBS for every second day. 3T3-L1 cells began to differentiate 1-2 days after the cells were put back in DMEM and 10% FBS. Between day 8-12 post-confluent, the cells became fully mature and these matured adipocytes will continue to store triglycerides. As shown in Fig. 2, the red area demonstrated the lipid droplets, stained with oil-red O.

Adipocytes play a central role in whole body energy homeostasis. Therefore, a lot of studies can be done by using these 3T3-L1 adipocytes such as morphological studies, glucose analysis, metabolome profiling, assessment of cell growth/proliferation, and assessment of cell viability and apoptosis.



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