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AML12 cells, also known as Alpha Mouse Liver 12 cells, are a non-tumorigenic epithelial cell line derived from the liver of a transgenic mouse. These cells were initially developed to provide a suitable in vitro model for studying the hepatocyte function and liver biology of the adult mouse. AML12 cells express characteristics typical of differentiated hepatocytes, including the production of albumin, transferrin, and other liver-specific proteins, making them an invaluable resource for research in toxicology, drug metabolism, and liver disease.
The cell line was established from hepatocytes isolated from a mouse harboring a transgene for human transforming growth factor alpha (TGF-alpha), under the control of the mouse metallothionein-I promoter. This genetic alteration contributes to the immortalization of the cells without disrupting their differentiated state. AML12 cells maintain a stable phenotype and karyotype under standard cell culture conditions, which includes a unique requirement for dexamethasone and insulin-transferrin-selenium in the growth medium to promote proliferation and maintain hepatocyte-specific functions.
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Products: The cells express high levels of human TGF alpha and lower levels of mouse TGF alpha.
- cultureMedium: DMEM:Ham's F12 (1:1), w: 3.1 g/L Glucose, w: 2.5 mM L-Glutamine, w: 15 mM HEPES, w: 0.5 mM Sodium pyruvate, w: 1.2 g/L NaHCO3 (Cytion article number 820400a)
- supplements: Supplement the medium with 10% FBS, 10 microgram/mL insulin, 5.5 microgram/mL transferrin, 5 ng/mL selenium, 40 ng/mL dexamethasone
- 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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