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NCI-H2452; Mesothelioma; Human (Homo sapiens) Item ID: ATCCRL5946

Permits and Restrictions

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Organism Homo sapiens, human
Tissue lung
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 1

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.
Disease mesothelioma
Gender male
Storage Conditions liquid nitrogen vapor phase
Derivation The line was established in November 1990 from a non-smoker with mesothelioma.
Clinical Data male The tissue donor was a non-smoker.
Comments The line was established in November 1990.
Complete Growth Medium The base medium for this cell line is ATCC-formulated RPMI-1640 Medium, ATCC 30-2001. To make the complete growth medium, add the following components to the base medium: fetal bovine serum (ATCC 30-2020) to a final concentration of 10%.
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.
  1. Remove and discard culture medium.
  2. 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.
  3. 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.
  4. Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
  5. Add appropriate aliquots of the cell suspension to new culture vessels.
  6. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 to 1:6 is recommended Medium Renewal: Every 2 to 3 days
Cryopreservation Freeze medium: Complete growth medium supplemented with 5% (v/v) DMSO Storage temperature: liquid nitrogen vapor phase
Culture Conditions Temperature: 37°C
STR Profile Amelogenin: X,Y CSF1PO: 11,12 D13S317: 12 D16S539: 11,13 D5S818: 11,12 D7S820: 9,11 THO1: 6,9.3 TPOX: 8,11 vWA: 17,18
Name of Depositor AF Gazdar, JD Minna
Year of Origin November, 1990
References

NCI-Navy Medical Oncology Branch Cell Line Supplement. J. Cell. Biochem. suppl. 24: 1996.

Sekido Y, et al. Neurofibromatosis type 2 (NF2) gene is somatically mutated in mesothelioma but not in lung cancer. Cancer Res. 55: 1227-1231, 1995. PubMed: 7882313

Penno MB, et al. High CD44 Expression on Human Mesotheliomas Mediates Association with Hyaluronan. Cancer J. Sci. Am. 1: 196-203, 1995.

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