HCE-2 [50.B1] (ATCC® CRL-11135)

Organism: Homo sapiens, human  /  Cell Type: epithelial Adenovirus 12-SV40 hybrid transformed  /  Tissue: eye, cornea  / 

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Organism Homo sapiens, human
Tissue
eye, cornea
Cell Type epithelial Adenovirus 12-SV40 hybrid transformed
Product Format frozen
Morphology epithelial
Culture Properties adherent
Biosafety Level 2 [Cells contain Adeno-12/SV-40 viral sequences]
Gender male
Ethnicity Black
Storage Conditions liquid nitrogen vapor phase
Karyotype aneuploid; Y chromosome is present
Images
Derivation
A primary culture of normal corneal epithelium was immortalized with Ad12-SV40 hybrid virus by overnight incubation with virus at 37°C
Clinical Data
Black
male
Effects
Yes, in soft agar
Comments
Keratinocyte Serum-Free medium is available from GIBCO, Grand Island, New York.
Complete Growth Medium Keratinocyte-Serum Free Medium (Gibco 17005-042)
Supplemented with frozen additives included (from GIBCO):
1) 0.05 mg/ml bovine pituitary extract (BPE)
2) 5 ng/ml epidermal growth factor (EGF).
NOTE: Do not filter EGF
And also supplemented with 500 ng/ml hydrocortisone and 0.005mg/ml insulin (not included).
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 0.05% (w/v) Trypsin-0.53mM EDTA solution.
  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 with 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. (Or neutralize with medium containing 10% fetal bovine serum).
  5. To remove trypsin-EDTA solution, transfer cell suspension to centrifuge tube and spin at approximately 125 x g for 5 to 10 minutes.
  6. Discard supernatant and resuspend cells in fresh growth medium. Add appropriate aliquots of cell suspension to new culture vessels. Seed cells on flasks precoated with a mixture of 0.01 mg/ml fibronectin, 0.03 mg/mL bovine collagen type I and 0.01 mg/mL bovine serum albumin.
  7. Incubate cultures at 37°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 is recommended
Medium Renewal: Twice per week
Cryopreservation
Freeze medium: 85% Growth Medium, 10% FBS, 5% DMSO
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Temperature: 37°C
STR Profile
Amelogenin: X,Y
CSF1PO: 10,13
D13S317: 8,11
D16S539: 9,11
D5S818: 11,12
D7S820: 8,10
THO1: 6,9
TPOX: 10,11
vWA: 15,18
Isoenzymes
AK-1, 1
ES-D, 1
G6PD, A
GLO-I, 2
Me-2, 1-2
PGM1, 1
PGM3, 2
Population Doubling Time 30 hrs
Name of Depositor CR Kahn, Gillette Medical Evaluation Laboratories
References

Kahn CR, Rhim J. Human corneal epithelial cell lines with extended lifespan. US Patent 5,585,265 dated Dec 17 1996

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

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
FAQ's
  1. Morphology of ATCC® CRL-11135


    Date Updated: 2/5/2013

  2. Slow growth for ATCC® CRL-11135


    Date Updated: 9/20/2013

References

Kahn CR, Rhim J. Human corneal epithelial cell lines with extended lifespan. US Patent 5,585,265 dated Dec 17 1996

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