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微信小章| 产品名称： | NIT-1 |
|---|---|
| 商品货号： | TS211225 |
| 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 xa0 xa0Cells contain polyomavirus DNA sequences
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| 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： | xad 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： | Hams F12K medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 90%; heat-inactivated dialyzed fetal bovine serum, 10%.
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| 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 25xa0cm2 flask (5 mL per 75xa0cm2 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 25xa0cm2 flask (10 mL per 75 cm2xa0flask) 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 |
| Deposited As： | mouse, transgenic for SV40 large T antigen |
| 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 |