NIT-1 (ATCC® CRL-2055)

Organism: Mus musculus, transgenic for SV40 large T antigen, mouse, transgenic for SV40 large T antigen  /  Cell Type: beta cell  /  Tissue: pancreas/islet of Langerhans  /  Disease: insulinoma

Organism Mus musculus, transgenic for SV40 large T antigen, mouse, transgenic for SV40 large T antigen
Tissue pancreas/islet of Langerhans
Cell Type beta cell
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2    [Cells contain polyomavirus DNA sequences]
Disease insulinoma
Age 10 weeks
Gender female
Strain NOD/Lt
Applications This cell line is a suitable transfection host.
Storage Conditions liquid nitrogen vapor phase
Derivation
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The NIT-1 cell line was derived from NOD/Lt mice.
Clinical Data
female
10 weeks
Antigen Expression
H-2 g7
Genes Expressed

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.

These cells express insulin

Comments 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.

Complete Growth Medium Ham's F12K medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 90%; heat-inactivated dialyzed fetal bovine serum, 10%.
Subculturing
Subcultures are prepared using a cell dissociation buffer (an enzyme free Hanks' based solution; Catalog number: 13150-016 available from GIBCO).
Remove the medium from the culture flask, add 2 mL of cell dissociation buffer per 25 cm2 flask (5 mL per 75 cm2 flask and gently rock the flask at room temperature for 1 to 2 minutes to bathe the cells in the buffer.
Aspirate the solution and discard. Allow the flask to sit at room temperature for 3 to 4 additional minutes (total time from initial addition of cell dissociation buffer is approximately 5 minutes).
Firmly tap the flask against the palm of the hand to dislodge cells.
Add 5 mL of fresh medium per 25 cm2 flask (10 mL per 75 cm2 flask) and triturate up and down directing the stream along the bottom of the flask to dislodge the cells and break up some of the clumps.
Add fresh medium, aspirate and dispense into new flasks.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:3 is recommended
Medium Renewal: 1 to 2 times per week
Cryopreservation
Culture medium, 95%; DMSO, 5%
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Name of Depositor EH Leiter
References

Hamaguchi K, et al. NIT-1, a pancreatic beta-cell line established from a transgenic NOD/Lt mouse. Diabetes 40: 842-849, 1991. PubMed: 1647994

Basic Documentation
References

Hamaguchi K, et al. NIT-1, a pancreatic beta-cell line established from a transgenic NOD/Lt mouse. Diabetes 40: 842-849, 1991. PubMed: 1647994