Immortalized Mouse Mammary Epithelial Cells (iMMEC)

The Immortalized Mouse Mammary Epithelial Cells (iMMEC) is derived by co-expression of adenovirus E1A and dominant-negative p53 (p53DD). This cell line is unique in that 1) it expresses surface markers, including estrogen-receptor alpha (ERα) and retains the functional properties characteristic to mammary epithelial cell and 2) upon lactogenic stimulation it forms dome-like structures indicative of mammary alveoli and secrets β-catenin into the acinar lumen. This cell line is suitable for studies in hormone regulation and 3D dimensional ductal morphogenesis.

Immortalization Method: Serial passaging and electroporation transfection with linearized DNA of E1A and p53DD

BioSafety Level: II

Species: Mouse

Source Organ: Mammary Epithelia

Growth Properties: Adherent

Morphology: Cuboidal

Population Doubling: 40 - 50 hours

Seeding Density: 25,000 - 40,000 cells/cm²

Markers: β-catenin, E-cadherin, Ep-CAM, CK8 and CK18; note that these cells lack drug-resistance markers

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow IV medium available at abm (TM004). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration on 10%, 5 µg/ml insulin (TM053), 1 µg/ml hydrocortisone, and 5 ng/ml EGF (Z200025).
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.

Lactogenic stimulation can be accomplished by inducing the cells with differentiation medium for 6 days (with media change every 2 days). To make complete differentiation medium, add the following components to the basal Prigorw IV medium (TM004): 5 µg/ml insulin (TM053), 1 µg/ml hydrocortisone, 3 µg/ml prolactin and 2% Matrigel.* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1. Western blot was used to confirm the presence of E1A and p53DD gene in the immortalized cell line. 2. Western blot and immunofluorescence were used to check gene expression of β-catenin, E-cadherin, Ep-CAM, CK8, CK18 and ERα.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C