AML12 (ATCC® CRL-2254)

Organism: Mus musculus, mouse  /  Cell Type: hepatocyte  /  Tissue: liver  /  Disease: normal

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Organism Mus musculus, mouse
Tissue liver
Cell Type hepatocyte
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1
Disease normal
Age 3 months
Gender male
Strain CD-1
Applications
This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Derivation
The AML12 (alpha mouse liver 12) cell line was established from hepatocytes from a mouse (CD1 strain, line MT42) transgenic for human TGF alpha.
Clinical Data
male
Genes Expressed
albumin; human transforming growth factor alpha (TGF alpha); mouse TGF alpha. 
Tumorigenic No
Effects
No, the cells do not form colonies or grow in soft agar.
No, the cells were not tumorigenic in immunosuppressed mice
Comments
The AML12 (alpha mouse liver 12) cell line was established from hepatocytes from a mouse (CD1 strain, line MT42) transgenic for human TGF alpha.
By electron microscopy, these cells exhibit typical hepatocyte features such as peroxisomes and bile canalicular like structure.
AML12 cells retain the capacity to express high levels of mRNA for serum (albumin, alpha 1 antitrypsin and transferrin) and gap junction (connexins 26 and 32) proteins, and contain solely isoenzyme 5 of lactate dehydrogenase.
The cells express high levels of human TGF alpha and lower levels of mouse TGF alpha.
Expression of liver specific proteins decreases with time in culture, but is reactivated by growing the cells in serum free medium.
Complete Growth Medium A 1:1 mixture of Dulbecco's modified Eagle's medium and Ham's F12 medium with 0.005 mg/ml insulin, 0.005 mg/ml transferrin, 5 ng/ml selenium, and 40 ng/ml dexamethasone, 90%; fetal bovine serum, 10%
Subculturing Volumes used in this protocol are for 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes.

  1. Remove and discard culture medium.
  2. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor.
  3. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes).
    Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal.
  4. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.

Subcultivation Ratio: 1:4 to 1:6
Medium Renewal: 2 to 3 times a week.
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.

Cryopreservation
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Population Doubling Time 37 hrs
Name of Depositor N Fausto
References

Wu JC, et al. Establishment and characterization of differentiated, nontransformed hepatocyte cell lines derived from mice transgenic for transforming growth factor alpha. Proc. Natl. Acad. Sci. USA 91: 674-678, 1994. PubMed: 7904757

Dumenco L, et al. Introduction of a murine p53 mutation corresponding to human codon 249 into a murine hepatocyte cell line results in growth advantage, but not in transformation. Hepatology 22: 1279-1288, 1995. PubMed: 7557882

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
References

Wu JC, et al. Establishment and characterization of differentiated, nontransformed hepatocyte cell lines derived from mice transgenic for transforming growth factor alpha. Proc. Natl. Acad. Sci. USA 91: 674-678, 1994. PubMed: 7904757

Dumenco L, et al. Introduction of a murine p53 mutation corresponding to human codon 249 into a murine hepatocyte cell line results in growth advantage, but not in transformation. Hepatology 22: 1279-1288, 1995. PubMed: 7557882