HIT-T15細胞

細胞類型： 其他細胞類型

細胞形態(tài)： 上皮樣

是否是腫瘤細胞： 0

物種來源： 其他

數(shù)量： 大量

器官來源： 胰腺

組織來源： islet of Langerhans

運輸方式： 凍存運輸

生長狀態(tài)： 貼壁生長

ATCC Number： CRL-1777?

Designations: HIT-T15

Depositors: R Santerre

HIT-T15細胞Biosafety Level: 2 [Cells contain polyomavirus DNA sequences ]

Shipped: frozen

Medium & Serum: See Propagation

Growth Properties: adherent

Organism: Mesocricetus auratus

Morphology: epithelial

Source: Organ: pancreas

Tissue: islet of Langerhans

Cell Type: beta cell;

Cellular Products: insulin

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Applications: transfection host

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).

HIT-T15細胞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%

Atmosphere: air, 95%; carbon dioxide (CO2), 5%

Temperature: 37.0°C

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: HIT-T15細胞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. HIT-T15細胞PubMed: 1634534