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Cell Biology ATCC® Number: CRL-1777™ Designations: HIT-T15 Price: Depositors: R Santerre Biosafety Level: 2 [Cells Contain PAPOVAVIRUS] Shipped: frozen Medium & Serum: See Propagation Growth Properties: adherent Organism: Mesocricetus auratus (hamster, Syrian golden) Morphology: epithelial Source: Organ: pancreas Tissue: islet of Langerhans Cell type: beta cell Cellular Products: insulin Permits/Forms: In addition to the MTA mentioned above, other ATCC and/or regulatory permits may be required for the transfer of this ATCC material. Anyone purchasing ATCC material is ultimately responsible for obtaining the permits. Please click here for information regarding the specific requirements for shipment to your location. Related Cell Culture Products Applications: transfection host (Roche FuGENE® Transfection Reagents) Receptors: glucagon; somatostatin; glucocorticoid Comments: This line was established from a primary culture of Syrian hamster islet cells which were transformed with SV40. The cells secrete up to 24 ng of insulin per 10 exp6 cells per 24 hours (passage 60). Insulin secretion is stimulated by glucose and glucagon, and is suppressed by somatostatin and glucocorticoids. Insulin synthesis decreases with length of time in culture. Propagation: ATCC complete growth medium: Ham's F12K medium with 2 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate, 87.5%; dialyzed horse serum, 10%; fetal bovine serum, 2.5% Temperature: 37.0C Atmosphere: air, 95%; carbon dioxide (CO2), 5% Subculturing: Protocol: Remove and discard culture medium. Briefly rinse the cell layer with 0.25% (w/v) Trypsin- 0.53 mM EDTA solution to remove all traces of serum that contains trypsin inhibitor. Add 2.0 to 3.0 ml of Trypsin-EDTA solution to flask and observe cells under an inverted microscope until cell layer is dispersed (usually within 5 to 15 minutes). Note: To avoid clumping do not agitate the cells by hitting or shaking the flask while waiting for the cells to detach. Cells that are difficult to detach may be placed at 37°C to facilitate dispersal. Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting. Add appropriate aliquots of the cell suspension to new culture vessels. Incubate cultures at 37°C. Interval: An inoculum of approximately 1.3 X 10(5) viable cells per sq. cm is recommended. Medium renewal: 2 to 3 times per week Preservation: Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase Related Products: Recommended medium (without the additional supplements or serum described under ATCC Medium): ATCC 30-2004 recommended serum: ATCC 30-2020 References: 22694: Lord JM , Ashcroft SJ . Identification and characterization of Ca2+-phospholipid-dependent protein kinase in rat islets and hamster beta-cells. Biochem. J. 219: 547-551, 1984. PubMed: 6234883 23275: Santerre RF , et al. Insulin synthesis in a clonal cell line of simian virus 40-transformed hamster pancreatic beta cells. Proc. Natl. Acad. Sci. USA 78: 4339-4343, 1981. PubMed: 6270673 23561: Aguilar-Bryan L , et al. Co-expression of sulfonylurea receptors and KATP channels in hamster insulinoma tumor (HIT) cells. Evidence for direct association of the receptor with the channel. J. Biol. Chem. 267: 14934-14940, 1992. PubMed: 1634534