01General information
- Description
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.
- Tissue
- Liver
- Applications
- 3D cell culture, High-throughput screening, Toxicology
- Synonyms
- AML-12
- AML 12
- Alpha Mouse Liver 12
02Classification
- Organism
- Mouse
- Sex
- Male
- Growth properties
- Adherent
- GMO status
- GMO-S1
- GMO status details
- GMO-S1: This murine hepatocyte cell line (AML12) contains a human TGF-α transgene introduced by transfection, enabling growth-factor-dependent signaling studies. The insert is stably integrated into hepatocytic cells. This classification applies only within Germany and may differ elsewhere.
- Biosafety level
- 1
03Characteristics
- Morphology
- Epithelial
- Cell type
- Hepatocyte
04Identifiers
- Citation
- AML12 (Cytion catalog number 300643)
- NCBI TaxID
- 10090
- Cellosaurus
- CVCL_0140
05Expression and susceptibility
- Products
- The cells express high levels of human TGF alpha and lower levels of mouse TGF alpha.
06Handling and culture
- Culture medium
- 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)
- Incubation atmosphere
37°C, 5% CO2, humidified atmosphere.
- Supplements
- Supplement the medium with 10% FBS, 10 microgram/mL insulin, 5.5 microgram/mL transferrin, 5 ng/mL selenium, 40 ng/mL dexamethasone
- Dissociation reagent
- 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.
- Thawing and culturing
Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.
Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.
For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.
Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.
Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.
Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.
Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.
Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.
07Quality control and cryopreservation
- Sterility
Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods.
To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections.
- Freeze medium
- As a cryopreservation medium, we 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.
- Storage conditions
For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.
- Shipping conditions
Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.
08Donor
- Breed / subspecies
- CD-1 MT42 transgenic
- Donor age
- 3 months
09Legal and safety
- Safety precautions
- When planning to store a cryovial in liquid nitrogen for future thawing, it is mandatory to adhere to stringent safety measures. Appropriate protective gloves and clothing are essential, and the use of a face mask or safety goggles is required during the transfer of frozen samples to or from the liquid nitrogen tank. This is to mitigate the risk of injury from potential cryovial explosions upon removal, which can result in the projection of sharp fragments.
- Warranty
At Cytion, we stand behind the quality and viability of our products. We guarantee that our cells will be viable at the start of culture for 60 days after shipment from our facilities. We assure that all materials conform to the specifications described in the Certificate of Analysis and/or Product Sheet.
For full details, please refer to our Warranty Statement.
- Material Transfer Agreement required
- Yes
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