tcel562 NIT-1

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Product Description

These mice are transgenic for the SV40 large T antigen under the control of a rat Insulin promoter, and spontaneously develop beta adenomas.At passage 18, most cells stained positively for Insulin, less than 5% were positive for glucagon and none were positive for somatostatin or pancreatic polypeptide.Insulin secretion is responsive to glucose concentration in the medium.There is low constitutive expression of MHC class I, class Ⅱ and ICAM-1 mRNA, but expression of both is markedly increased by treatment with interferon gamma.Stimulation of class I mRNA is accompanied by increased class I antigen expression and induction of an occult class I product expressing the H-2.39 specificity.MHC class Ⅱ antigen is not induced despite the induction of the mRNA.NIT-1 cells show ultrastructural features of differentiated mouse beta cells(well developed rough endoplasmic reticulum, extensive golgi apparatus and beta granules).The cells shed a mature ecotropic type C retrovirus. The retrovirus is capable of infecting other Fv-1 n mouse cell lines, so care should be taken to avoid cross infection. NOTE: NIT-1 cells will not form a confluent monolayer; however, they will form nice colonies of monolayered cells in a fairly dense array.When the NIT-1 colonies begin to ball up slightly and show many round cells on top of the monolayers as well as floating in the media, it is time to passage them.

Information

Applicationtransfection
TissueTransgenic for sv40 large t antigen pancreas/islet of langerhans
Morphologyepithelial
Growth Propertiesadherent

Specifications

Complete Growth MediumHam's F-12K (PM150910)+10% heat-inactivated dialyzed FBS (164210-500)+1% P/S (PB180120)
Subcultivation Ratio1:2-1:4
Medium Renewalevery 2 to 3 days
CryopreservationFreeze medium: 60% Basal medium+30% FBS+10% DMSO Storage temperature: Liquid nitrogen vapor phase
Culture ConditionsAtmosphere: Air, 95%; CO2, 5% Temperature: 37℃
Antigen ExpressionH-2 g7
Gene ExpressionThere is low constitutive expression of MHC class I, class Ⅱ and ICAM-1 mRNA, but expression of both is markedly increased by treatment with interferon gamma.These cells express Insulin.
Durationtransfection

Misc Information

SubculturingRemove and discard culture medium. Briefly rinse the cell layer with DPBS solution to remove all traces of serum that contains trypsin inhibitor. Add 1.0 to 2.0 mL of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 2 to 3 minutes). Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 4.0 to 6.0 mL of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels.
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