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FL 62891
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ImClone Systems Inc.
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Biosafety Level:
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2 [Cells contain SV-40 viral DNA sequences ]
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frozen
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See Propagation
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adherent
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Homo sapiens (human)
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fibroblast
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Organ: liver
Cell Type: immortalized with SV40 large T antigen
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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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fetus
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ATCC complete growth medium: Iscove's modified Dulbecco's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate supplemented with 0.1 mM non-essential amino acids, 1.0 mM sodium pyruvate, and 15% fetal bovine serum
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 32.0°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 which 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 32°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 32°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 is recommended
Medium Renewal: Every 2 to 3 days
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Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
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48 hours
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Recommended medium (without the additional supplements or serum described under ATCC Medium):ATCC 30-2005
recommended serum:ATCC 30-2020
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70742: Lemischka IR. Nucleic acids encoding fragments of hematopoietic stem cell receptor flk-2. US Patent 5,270,458 dated Dec 14 1993
70744: Lemischka IR. Tyrosine kinase receptor flk-2 and fragments thereof. US Patent 5,367,057 dated Nov 22 1994
70745: Lemischka IR. Tyrosine kinase receptor human flk-2-specific antibodies. US Patent 5,548,065 dated Aug 20 1996
70746: Lemischka IR. Nucleic acids encoding soluble human FLK-2 extracellular domain. US Patent 5,621,090 dated Apr 15 1997
70747: Lemischka IR. Antibodies against tyrosine kinase receptor flk-1. US Patent 5,747,651 dated May 5 1998
70748: Lemischka IR. Method for isolating stem cells expressing flk-1 receptors. US Patent 5,912,133 dated Jun 15 1999
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