HP [HT1080 poly] (ATCC® CRL-12012)

Organism: Homo sapiens, human  /  Disease: fibrosarcoma

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Organism Homo sapiens, human
Product Format frozen
Culture Properties adherent
Biosafety Level 2 Cells contain CMV viral DNA sequences
Disease fibrosarcoma
Age 35 years
Gender male
Ethnicity Caucasian
Applications
The HP polytropic retroviral packaging cell line was derived from the human fibrosarcoma cell line, HT-1080 (see ATCC CCL-121).
After selection for transfected cells, individual drug resistant cell colonies were expanded, analyzed and selected for overexpression of MoMLV gag/pol.
Storage Conditions liquid nitrogen vapor phase
Derivation
The HP polytropic retroviral packaging cell line was derived from the human fibrosarcoma cell line, HT-1080 (see ATCC CCL-121).
Clinical Data
male
Caucasian
35 years
Comments
The HP polytropic retroviral packaging cell line was derived from the human fibrosarcoma cell line, HT-1080 (see ATCC CCL-121).
HT1080 cells were co-transfected with the methotrexate resistance vector, pFR400 and the MoMLV gag/pol expression vector, pSCV10.
After selection for transfected cells, individual drug resistant cell colonies were expanded, analyzed and selected for overexpression of MoMLV gag/pol.
HT1080 (clone SCV21) was co-transfected with the phleomycin resistance vector, pUT507 and the polytropic envelope expression vector, pCMVMCF and subsequently cloned.
The HP clone was selected for expression of relatively high levels of both gag/pol and polytropic envelope proteins.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Dulbecco's Modified Eagle's Medium, Catalog No. 30-2002. To make the complete growth medium, add the following components to the base medium:
  • O.1 mM Non-Essential Amino Acids (NEAA)
  • fetal bovine serum to a final concentration of 10%

Subculturing
Protocol:
  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 1:8 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
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37.0°C
STR Profile
Amelogenin: X,Y
CSF1PO: 12
D13S317: 12,14
D16S539: 9,12
D5S818: 11,13
D7S820: 9,10
THO1: 6
TPOX: 8
vWA: 14,19
Name of Depositor Chiron Viagene, Inc.
References

Barber JR, et al. Retroviral packaging cell lines. US Patent 5,591,624 dated Jan 7 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
References

Barber JR, et al. Retroviral packaging cell lines. US Patent 5,591,624 dated Jan 7 1997