Immortalized Mouse Colonic Epithelial Cells-Conditionally (YAMC)

YAMC is a conditionally immortalized mouse colon epithelial cell line isolated from the mouse harboring thermolabile mutation (tsA58) under the control of an interferon (IFN)-γ-inducible H-2K^b promoter and a temperature-sensitive simian virus 40 large T antigen.
At a non-permissive temperature (37°C-39°C), the cells cease to proliferate and begin to detach from the plate. Cells in culture display some microvilli formation and expression levels of the brush border-associated enzymes, alkaline phosphatase, dipeptidyl peptidase IV, and sucrase, can be induced in the line by the addition of 1mM sodium butyrate, 1 µM Phorbol Myristate Acetate (PMA), or 1 µM Retanoic Acid (RA) to the culture medium for 5 days. Conversely, the addition of 5 µg/ml TGF-β or 1 µM Bt2GMP will downregulate the two enzymes.

Immortalization Method: Isolated from transgenic mouse carrying a temperature-sensitive simian virus 40 tumor antigen (tsA58)

BioSafety Level: II

Species: Mouse

Source Organ: Colon

Growth Properties: Adherent

Morphology: Epithelial

Population Doubling: 22 - 32 hours

Seeding Density: 20,000 cells/cm². Recommended split ratio of 1:3

Markers: CK18

Donor Age: N/A

Donor Gender: N/A

Donor Ethnicity: N/A

Propagation: The base medium for this cell line is Prigrow II medium available at abm (TM002). To make the completed growth medium, add the following components to the base medium: fetal bovine serum (TM999)* to a final concentration of 10%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1%, and Recombinant Murine IFNG (Z200085) to a final concentration of 5 ng/mL.
The expression of alkaline phosphatase and dipeptidyl peptidase IV can be upregulated by the addition of 1mM sodium butyrate, 1 µM Phorbol Myristate Acetate (PMA), or 1 µM Retinoic Acid (RA) to the culture medium for 5 days. Conversely, the addition of 5 µg/ml TGF-β or 1 µM Bt₂GMP will downregulate the two enzymes.
Carbon dioxide (CO₂): 5%, Temperature: 33.0°C.
* abm does not recommend to use heat-inactivated FBS for cell culture unless specified otherwise.

Quality Control: 1) Immunohistochemical staining was used to detect the presence of SV40 large T antigen and keratin 18; 2) Biochemical assays (i.e. method of Nagatsu et al and method of Dahlquist) were used to measure brush border-associated enzymes alkaline phosphatase, dipeptidyl peptidase IV and sucrase.

Shipping Condition: Dry Ice

Storage Condition: liquid nitrogen or -180°C