FL 62891 (ATCC® CRL-11005)

Organism: Homo sapiens, human  /  Cell Type: immortalized with SV40 large T antigen  / 

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Organism Homo sapiens, human
Cell Type immortalized with SV40 large T antigen
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 2 Cells contain SV-40 viral DNA sequences
Age fetus
Applications
The cells produce flk-1 and some flk-2 ligands.
The cell line was derived from human fetal liver tissue, and immortalized by transformation with an ecotropic retrovirus containing a temperature sensitive SV40 T-antigen (tsA58) and a G418 resistance gene.
Storage Conditions liquid nitrogen vapor phase
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Derivation
The cell line was derived from human fetal liver tissue, and immortalized by transformation with an ecotropic retrovirus containing a temperature sensitive SV40 T-antigen (tsA58) and a G418 resistance gene. After several days at 37C, the temperature of the medium was lowered to 32C. Cells were selected with G418. The selected cells were expanded and maintained. The cells produce flk-1 and some flk-2 ligands.
Comments
The cell line was derived from human fetal liver tissue, and immortalized by transformation with an ecotropic retrovirus containing a temperature sensitive SV40 T-antigen (tsA58) and a G418 resistance gene. After several days at 37C, the temperature of the medium was lowered to 32C. Cells were selected with G418. The selected cells were expanded and maintained. The cells produce flk-1 and some flk-2 ligands.
Complete Growth Medium Iscove's modified Dulbecco's medium with 4 mM L-glutamine adjusted to contain 1.5 g/L sodium bicarbonate supplemented with 0.1 mM non-essential amino acids, 1.0 mM sodium pyruvate, and 15% fetal bovine serum
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 which 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 32°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 32°C.
Subcultivation Ratio: A subcultivation ratio of 1:3 to 1:4 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: 32.0°C
Population Doubling Time 48 hours
Name of Depositor ImClone Systems Inc.
References

Lemischka IR. Nucleic acids encoding fragments of hematopoietic stem cell receptor flk-2. US Patent 5,270,458 dated Dec 14 1993

Lemischka IR. Tyrosine kinase receptor flk-2 and fragments thereof. US Patent 5,367,057 dated Nov 22 1994

Lemischka IR. Tyrosine kinase receptor human flk-2-specific antibodies. US Patent 5,548,065 dated Aug 20 1996

Lemischka IR. Nucleic acids encoding soluble human FLK-2 extracellular domain. US Patent 5,621,090 dated Apr 15 1997

Lemischka IR. Antibodies against tyrosine kinase receptor flk-1. US Patent 5,747,651 dated May 5 1998

Lemischka IR. Method for isolating stem cells expressing flk-1 receptors. US Patent 5,912,133 dated Jun 15 1999

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

Lemischka IR. Nucleic acids encoding fragments of hematopoietic stem cell receptor flk-2. US Patent 5,270,458 dated Dec 14 1993

Lemischka IR. Tyrosine kinase receptor flk-2 and fragments thereof. US Patent 5,367,057 dated Nov 22 1994

Lemischka IR. Tyrosine kinase receptor human flk-2-specific antibodies. US Patent 5,548,065 dated Aug 20 1996

Lemischka IR. Nucleic acids encoding soluble human FLK-2 extracellular domain. US Patent 5,621,090 dated Apr 15 1997

Lemischka IR. Antibodies against tyrosine kinase receptor flk-1. US Patent 5,747,651 dated May 5 1998

Lemischka IR. Method for isolating stem cells expressing flk-1 receptors. US Patent 5,912,133 dated Jun 15 1999