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Immortalized-(Conditionally)-Mouse-Small-Intestinal-Epithelial-Cells-(MSIE)

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Immortalized-(Conditionally)-Mouse-Small-Intestinal-Epithelial-Cells-(MSIE)

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Immortalized-(Conditionally)-Mouse-Small-Intestinal-Epithelial-Cells-(MSIE)

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	Print Version
BioSafety Level	II
Organism	18-day-old H-2K^b-tsA58 transgenic mouse
Source Organ	Small intestine
Growth Properties	Adherent
Recommended Seeding Density	Recommended split ratio of 1:3
Markers	CK18
Applications	For Research Use Only
Immortalization Method	Isolated from transgenic mouse carrying a temperature-sensitive simian virus 40 tumor antigen (tsA58)
Description	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.
Procedure Overview
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Propagation	Use of PriCoat? T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels with the following conditions. The base medium for this cell line is Prigrow II medium available from ABM (TM002). To make the completed growth medium, add the following components to the base medium: 5% fetal bovine serum (TM999), 5 U/ml murine IFN-γ and 1% Penicillin/Streptomycin (G255). Atmosphere: air: 95%, CO?: 5%; Temperature: 33.0°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.
Preservation	1. Freeze Medium: Complete growth medium with 20% FBS and 10% DMSO. 2. Storage Temperature: Liquid nitrogen vapour phase.
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.
Disclaimer	1. The CoA for this product (provided upon request) verifies the cell-type specific gene expression via RT-PCR only. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 2. We strongly recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 3. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.