Mouse Cell Lines-Cell Lines-InCellGene LLC.-CellLines,Biomedicine reasearch Product,Cell Biology,Cell Culture,293T Cell,Media

Organism	Mus musculus, mouse
Tissue	liver
Cell Type	hepatocyte
Product Format	frozen
Morphology	epithelial
Culture Properties	adherent
Biosafety Level	1 Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.
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.
Cellular Products	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	The base medium for this cell line is DMEM:F12 Medium . To make the complete growth medium, add the following component to the 500 mL of the base medium: Supplemented with: 10% fetal bovine serum (FBS) 10 μg/ml insulin 5.5 μg/ml transferrin 5 ng/ml selenium 40 ng/ml dexamethasone This medium is formulated for use with a 5% CO2 in air atmosphere.
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. Remove and discard culture medium. 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. 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. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. 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