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HIT-T15
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R Santerre
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Biosafety Level:
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2 [Cells Contain PAPOVAVIRUS ]
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frozen
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Medium & Serum:
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See Propagation
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adherent
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Mesocricetus auratus (hamster, Syrian golden)
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epithelial
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Organ: pancreas
Tissue: islet of Langerhans
Cell Type: beta cell;
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insulin
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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.
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transfection host (Roche FuGENE® Transfection Reagents)
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glucagon; somatostatin; glucocorticoid
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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
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Protocol:
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Remove and discard culture medium.
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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.
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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.
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Add 6.0 to 8.0 ml of complete growth medium and aspirate cells by gently pipetting.
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Add appropriate aliquots of the cell suspension to new culture vessels.
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Incubate cultures at 37°C.
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