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3T3-L1 cells are a clonal line of preadipocytes derived from mouse embryonic fibroblasts. These cells have become a widely used in vitro model for studying the process of adipogenesis, including adipogenesis and lipogenesis, which is the differentiation of preadipocytes into adipocytes (fat cells). The name "3T3" refers to the transfer (T) protocol that involved transferring the cells every 3 days, and "L1" signifies the particular clone that was isolated.
Initially, 3T3-L1 cells exhibit a fibroblast-like morphology, but upon induction of 3T3-L1 cell differentiation, 3T3-L1 cells change from a preadipocyte to a mature adipocyte state and accumulate lipid droplets, a hallmark of obesity and metabolic syndrome. The differentiation process from 3T3-L1 preadipocytes to 3T3-L1 adipocytes is triggered by a specific cocktail of inducers, typically including dexamethasone, 3-isobutyl-1-methylxanthine (IBMX), and insulin.
As 3T3-L1 adipocytes adopt the characteristics of mature adipocytes, they begin to express genes that are crucial for adipocyte function, such as those coding for enzymes involved in fatty acid metabolism and hormones like leptin and adiponectin, which play vital roles in regulating appetite, energy balance, and insulin sensitivity. Studying 3T3-L1 cell transformations enhances our understanding of adipogenesis and obesity and fat-related diseases, such as type 2 diabetes, by revealing how lipid accumulation in adipocytes leads to cellular dysfunction and broader metabolic issues.
Moreover, the 3T3-L1 cell line is instrumental in investigating the impact of various substances on adipocyte behavior, such as the effect of pharmacological agents on lipolysis or the anti-inflammatory properties of certain diets that may prevent insulin resistance.
3T3-L1 cells have been extensively used to study the molecular and cellular mechanisms underlying adipocyte differentiation, insulin sensitivity, lipid metabolism, and the effects of various nutritional and pharmacological agents on these processes. Given their ability to differentiate into adipocytes and their ease of culture in vitro, 3T3-L1 cells provide a valuable model system for obesity and diabetes research, as well as for the discovery of new therapeutic targets related to metabolic dise
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- Tumorigenic: No
- Virus susceptibility: murine leukemia virus, murine sarcoma virus, vesicular stomatitis, vaccinia, herpes simplex, N-tropic oncornaviruses C
- Products: Insulin, collagen, triglycerides
- Karyotype: 2n=40
- cultureMedium: DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
- supplements: Supplement the medium with 10% FBS
- dissociationReagent: Accutase
- subculturing: Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium.
- freezeMedium: As a cryopreservation medium, use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
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