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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
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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