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ATCC Number：CRL-2254?
相關疾病：正常
運輸方式：凍存運輸
細胞類型：其他細胞類型
是否是腫瘤細胞：0
物種來源：小鼠
數量：大量
品系：CD-1
生長狀態(tài)：貼壁生長
細胞形態(tài)：上皮樣
器官來源：肝
年限：3 months
規(guī)格：0.5mg Designations: AML12
Depositors: ?N Fausto
Biosafety Level:1
Shipped: frozen
Medium & Serum: See Propagation Growth Properties:adherent
Organism: Mus musculus
Morphology:epithelial


Source: Organ: liver
Strain: CD-1
Disease: normal
Cell Type: hepatocyte;
Cellular Products:albumin; human transforming growth factor alpha (TGF alpha); mouse TGF alpha
Permits/Forms:In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location.
Applications:transfection host (Roche Transfection Reagents )
Tumorigenic:No
Age: 3 months
Gender: male
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.
Propagation: ATCC 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%
Temperature: 37.0°C
Subculturing: Protocol:

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: A subcultivation ratio of 1:4 is recommended
   Medium Renewal: Every 2 to 3 days

Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Doubling Time: 37 hrs
Related Products:Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2006
recommended serum:ATCC 30-2020
References: 22600: 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
23390: 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