產(chǎn)品名稱 |
HIT-T15 |
商品貨號(hào) |
B164614 |
Organism |
Mesocricetus auratus, hamster, Syrian golden |
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]
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. |
Applications |
This cell line is a suitable transfection host. |
Storage Conditions |
liquid nitrogen vapor phase |
Images |
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Derivation |
This line was established from a primary culture of Syrian hamster islet cells which were transformed with SV40. |
Receptor Expression |
glucagon; somatostatin; glucocorticoid |
Genes Expressed |
insulin |
Comments |
The cells secrete up to 24 ng of insulin per 106 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. |
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%
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Subculturing |
Volumes are given for a 75 cm2 flask. Increase or decrease the amount of dissociation medium needed proportionally for culture vessels of other sizes.
- 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 105 viable cells per cm2 is recommended.
Medium Renewal: 2 to 3 times per week |
Cryopreservation |
Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase |
Culture Conditions |
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C |
Name of Depositor |
R Santerre |
Deposited As |
Mesocricetus auratus |
References |
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
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
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
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Cross References |
Nucleotide (GenBank) :
U62810
Mesocricetus auratus potassium channel Kv8.1 mRNA, complete cds.
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