微信小陈
微信小章| 产品名称： | NIT-2 |
|---|---|
| 商品货号： | TS211227 |
| Organism： | Mus musculus, transgenic for SV40 large T antigen, mouse, transgenic for SV40 large T antigen |
| Tissue： | pancreas |
| Cell Type： | beta cell |
| Product Format： | frozen |
| Morphology： | epithelial |
| Culture Properties： | adherent |
| Biosafety Level： | 2 Cells 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： | adenoma; carboxypeptidase E defective |
| Age： | 10 week old |
| Gender： | male |
| Applications： | The NIT-2 cell line was derived from the pancreatic beta cells of Cpe(fat)/Cpe(fat) mice by crossing C57BLKS/J-Cpe(fat)/+ mice with NOD/Lt-Tg(RIPTag)1Lt mice. NOD/Lt-Tg(RIPTag)1Lt mice are transgenic for the SV40 large T antigen under the control of a rat insulin promoter, and spontaneously develop beta adenomas. Pro-CPE, but not the mature form of CPE, is present in NIT-2 cells, and neither pro-CPE nor CPE are secreted into the medium. The secretion of insulin/proinsulin from NIT-2 cells is significantly elevated by secretagogues, indicating that CPE is not required for sorting proinsulin into the regulated pathway. |
| Storage Conditions： | liquid nitorgen vapor phase |
| Derivation： | The NIT-2 cell line was derived from the pancreatic beta cells of Cpe(fat)/Cpe(fat) mice by crossing C57BLKS/J-Cpe(fat)/+ mice with NOD/Lt-Tg(RIPTag)1Lt mice. The NIT-2 cell line was cultured from adenomatous islets obtained from a 10 week old F2 male and was compared with the NIT-1 cell line (ATCC-CRL-2055) previously developed from mice with wild-type CPE. |
| Clinical Data： | The NIT-2 cell line was cultured from adenomatous islets obtained from a 10 week old F2 male and was compared with the NIT-1 cell line (ATCC-CRL-2055) previously developed from mice with wild-type CPE. male |
| Comments： | The NIT-2 cell line was derived from the pancreatic beta cells of Cpe(fat)/Cpe(fat) mice by crossing C57BLKS/J-Cpe(fat)/+ mice with NOD/Lt-Tg(RIPTag)1Lt mice. NOD/Lt-Tg(RIPTag)1Lt mice are transgenic for the SV40 large T antigen under the control of a rat insulin promoter, and spontaneously develop beta adenomas. Carboxypeptidase E is required for complete conversion of proinsulin to mature insulin. A spontaneous point mutation in the coding region of the carboxypeptidase E (CPE) gene in Cpe(fat)/Cpe(fat) mice affects proinsulin processing. The NIT-2 cell line was cultured from adenomatous islets obtained from a 10 week old F2 male and was compared with the NIT-1 cell line (ATCC-CRL-2055) previously developed from mice with wild-type CPE. Electron microscopy of the cultured NIT-2 showed increased numbers of enlarged and electron-lucent granules compared with NIT-1 cells. Pro-CPE, but not the mature form of CPE, is present in NIT-2 cells, and neither pro-CPE nor CPE are secreted into the medium. Proinsulin is less extensively processed in NIT-2 than in NIT-1 cells, indicating that the Cpe(fat)mutation affects both the endopeptidase and carboxypeptidase reactions. The secretion of insulin/proinsulin from NIT-2 cells is significantly elevated by secretagogues, indicating that CPE is not required for sorting proinsulin into the regulated pathway. |
| 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%.
|
| Subculturing： | Volumes used in this protocol are for 25 cm2 or 75 cm2 flask; proportionally reduce or increase amount of dissociation medium for culture vessels of other sizes. NOTE: NIT-2 cells will not form a confluent monolayer, however, they will form nice colonies of monolayered cells in a fairly dense array. When the NIT-2 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.
Subcultivation Ratio: 1:2 to 1:3 Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 10 in Culture of Animal Cells, a manual of Basic Technique by R. Ian Freshney, 3rd edition, published by Alan R. Liss, N.Y., 1994.xa0xa0 |
| Cryopreservation： | Complete growth medium described above supplemented with 5% (v/v) DMSO.xa0 Cell culture tested DMSO is available as ATCC Catalog No. 4-X. |
| 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： | Varlamov O, et al. Beta-cell lines derived from transgenic Cpe(fat)/Cpe(fat) mice are defective in carboxypeptidase E and proinsulin processing. Endocrinology 138: 4883-4892, 1997. PubMed: 9348219 Naggert JK, et al. Hyperproinsulinaemia in obese fat/fat mice associated with a carboxypeptidase E mutation which reduces enzyme activity. Nat. Genet. 10: 135-142, 1995. PubMed: 7663508 Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC. Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988. Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC. |