Immortalized Mouse Small Intestinal Epithelial Cells-Conditionally (MSIE)

MSIE is a conditionally immortalized mouse small intestine 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. The functional expression of the SV40 large T antigen is induced by culturing the cells in vitro in medium containing IFN γ at a temperature permissive (33°C). At a non-permissive temperature (37°C-39°C), the cells cease to proliferate and shows a 2-3X differentiation rate as indicated by expression levels of the brush border-associated enzymes, alkaline phosphatase, dipeptidyl peptidase IV, and sucrase.

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

BioSafety Level: II

Species: Mouse

Source Organ: Small intestine

Growth Properties: Adherent

Morphology: Other

Population Doubling: N/A

Seeding Density: 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 5%, 5 U/ml murine IFN-γ and Penicillin/Streptomycin Solution (G255) to a final concentration of 1%
Carbon dioxide (CO₂): 5%, Temperature: +37.0°C.
The expression of alkaline phosphatase and dipeptidyl peptidase IV can be upregulated by the addition of 1mM sodium butyrate, 1 µM PMA, or 1 µM 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.
* 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