Immortalized Mouse Colonic Epithelial Cells-Conditionally (YAMC) | T0567

Immortalized Mouse Colonic Epithelial Cells-Conditionally (YAMC) | T0567

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

Organism:

18-day-old H-2K^b-tsA58 transgenic mouse

Species:

Mouse

Source Organ:

Colon

Organ Type:

Digestive

Growth Properties:

Adherent

Morphology:

Epithelial

Passage Number:

N/A

Population Doubling:

22 - 32 hours

Seeding Density:

20,000 cells/cm10². 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 . To make the completed growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%, Penicillin/Streptomycin Solution to a final concentration of 1%, and Recombinant Murine IFNG 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 -180C