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MeT-5A
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The United States of America
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Biosafety Level:
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2 [Cells contain polyomavirus DNA sequences ]
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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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Homo sapiens (human)
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epithelial
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Tissue: mesothelium
Cell Type: epithelialvirus transformed
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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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No
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Amelogenin: X,Y
CSF1PO: 10,12
D13S317: 11,13
D16S539: 12
D5S818: 12
D7S820: 10
THO1: 6,9.3
TPOX: 8
vWA: 15,18
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ATCC complete growth medium: Medium199 with Earle's BSS, .75 mM L-glutamine and 1.25 g/L sodium bicarbonate supplemented with with 3.3 nM epidermal growth factor (EGF), 400 nM hydrocortisone, 870 nM insulin, 20 mM HEPES, trace elements (see above reference) and 10% fetal bovine serum.
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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To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 xg for 5 to 10 minutes.Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels.
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Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:2 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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recommended serum:ATCC 30-2020
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21937: Reddel RR, et al. Immortalized human bronchial epitherial mesothelial cell lines. US Patent 4,885,238 dated Dec 5 1989
23292: Lechner JF, et al. Asbestos-associated chromosomal changes in human mesothelial cells. Proc. Natl. Acad. Sci. USA 82: 3884-3888, 1985. PubMed: 2987952
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