10.014 pRSV-T (ATCC® CRL-11515)

Organism: Homo sapiens, human  /  Cell Type: epithelial cells transfected with a plasmid containing the SV40 early region  /  Tissue: eye, cornea  / 

Permits and Restrictions

View Permits

Organism Homo sapiens, human
Tissue
eye, cornea
Cell Type epithelial cells transfected with a plasmid containing the SV40 early region
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain SV-40 and Rous Sarcoma viral sequences]
Storage Conditions liquid nitrogen vapor phase
Images
Derivation
A primary culture of normal corneal epithelium was immortalized by transfection with plasmid pRSV-T using lipofectamine overnight at 37°C. pRSV-T contains the SV40 early region genes and the Rous Sarcoma virus long terminal repeat.
Complete Growth Medium Keratinocyte-Serum Free medium (GIBCO-BRL 17005-042) with 5 ng/ml human recombinant EGF, 0.05 mg/ml bovine pituitary extract, 0.005 mg/ml insulin and 500 ng/ml hydrocortisone.
Subculturing
Subculture when 80% confluent
  1. remove medium
  2. briefly rinse the cell layer with 0.05% (w/v) Trypsin-53mM EDTA solution
  3. add Trypsin-EDTA solution to flask and incubate at 37°C for up to three minutes to facilitate dispersal. Observe cells under an inverted microscope. Hit or shake the flask if some cells remain attached
  4. neutralize the trypsin with growth medium, centrifuge at 300 x g for 8 minutes
  5. resuspend the pellet in fresh growth medium and dispense into new COATED flasks. Flasks used to propagate these cells must be coated (for at least 2 hours at 37°C) with 0.01 mg/mL bovine serum albumin, 0.01 mg/mL fibronectin and 0.03 mg/mL bovine collagen type I.
  6. Incubate at 37°C with 5% CO2. Cells will not be well attached for the first 24 hours after subculture.
Subcultivation Ratio: A subcultivation ratio of 1:2 to 1:4 is recommended when 80% confluent
Medium Renewal: Every 2 to 3 days
Cryopreservation
Freeze medium: complete growth medium supplemented with 7.5% (v/v) DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
STR Profile
Amelogenin: X,Y
CSF1PO: 10
D13S317: 12,13
D16S539: 8,13
D5S818: 12,13
D7S820: 8,10
THO1: 6,8
TPOX: 8,10
vWA: 15
Name of Depositor CR Kahn, Gillette Medical Evaluation Laboratories
References

Walker TL, Kahn CR. Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997

Kruszewski FH, et al. Evaluation of a human corneal epithelial cell line as an in vitro model for assessing ocular irritation. Fundam. Appl. Toxicol. 36: 130-140, 1997. PubMed: 9143482

Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
References

Walker TL, Kahn CR. Human corneal epithelial cell lines with extended lifespan. US Patent 5,672,498 dated Sep 30 1997

Kruszewski FH, et al. Evaluation of a human corneal epithelial cell line as an in vitro model for assessing ocular irritation. Fundam. Appl. Toxicol. 36: 130-140, 1997. PubMed: 9143482