BJ-5ta (ATCC® CRL-4001)

Organism: Homo sapiens, human  /  Cell Type: Fibroblasts immortalized with hTERT  /  Tissue: Foreskin  /  Disease: Normal

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Organism Homo sapiens, human
Tissue Foreskin
Cell Type Fibroblasts immortalized with hTERT
Product Format frozen
Morphology Fibroblast-like
Culture Properties Adherent
Biosafety Level 1
Disease Normal
Age Neonatal
Gender Male
Storage Conditions Liquid nitrogen vapor phase
Karyotype This is a diploid human cell line of male origin with a modal chromosome number of 46 that occurred in 90% of the cells counted. The sex chromosomes, X and Y are both karyotypically normal.
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Derivation

The hTERT-immortalized foreskin fibroblast cell line, BJ-5ta, was derived by transfecting the BJ foreskin fibroblast cell line with the pGRN145 hTERT-expressing plasmid (ATCC MBA-141) at population doubling 58. Cells were cultured in medium containing hygromycin B until stable clones were selected [Pubmed: 9454332]. 

Antigen Expression

Antigen expression: Positive for fibroblast surface protein; Homo sapiens, expressed (fibroblast surface protein (FSP) was assayed by flow cytometry.).

Negative for the pan-cytokeratin epithelial marker; Homo sapiens (cytokeratins were assayed by immunocytochemistry using a pan-cytokeratin antibody).

Complete Growth Medium A 4:1 mixture of Dulbecco's medium and Medium 199 with supplements as follows :
4 parts of Dulbecco's Modified Eagle's Medium containing 4 mM L-glutamine, 4.5 g/L glucose and 1.5 g/L sodium bicarbonate
1 part of Medium 199
Supplemented with:
0.01 mg/ml hygromycin B
10% fetal bovine serum
Subculturing
Volumes are given for a 75 cm2 flask; proportionally reduce or increase the amount of dissociation medium for culture vessels of other sizes.
  1. Remove and discard culture medium.
  2. Add 3.0 to 5.0 mL of 0.25% trypsin-0.53 mM EDTA solution to the flask and observe cells under an inverted microscope until the cell layer is dispersed (usually within 5 to 15 minutes).
  3. Note: To avoid clumping do not hit or shake 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 10.0 mL of complete growth medium and aspirate cells by gently pipetting. 
  5. Add appropriate aliquots of the cell suspension to new culture vessels. An inoculum of 3 x 103 to 5 x 103 viable cells/cm2 is recommended. Maintain cultures at a cell concentration between 8 x 103 and 1 x 104 cells/cm2.
  6. Subcultivation ratio: 1:2 to 1:3 twice weekly
  7. Incubate cultures at 37°C.
Subcultivation Ratio: 1:2 to 1:3 twice weekly
Medium Renewal: every 2 to 3 days
Note: For more information on enzymatic dissociation and subculturing of cell lines consult Chapter 13 in Culture Of Animal Cells: A Manual of Basic Technique by R. Ian Freshney, 5th edition, published by Wiley-Liss, N.Y., 2005.
Cryopreservation
Freeze medium: culture medium, 30%; fetal bovine serum, 60%; DMSO, 10%
Storage temperature: liquid nitrogen vapor phase
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Growth Conditions: Subculture when cell concentration reaches between 8 X 103 and 1 X 104 cells/cm2.
STR Profile
Amelogenin: X,Y
CSF1PO: 10,12
D13S317: 8,9
D16S539: 9,13
D5S818: 12
D7S820: 11,12
THO1: 7,8
TPOX: 10,11
vWA: 16,18
Population Doubling Level (PDL) As part of our quality control, we have tested this cell line for its ability to grow for a minimum of 15 population doublings after recovery from cryopreservation. We have also compared its karyotype, telomerase expression level, growth rate, morphology and tissue-specific markers when first recovered from cryopreservation with that of cells at 10+ population doublings to ensure that there is no change in these parameters and that the cells are capable of extended proliferation.
Name of Depositor Geron Corporation
References

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

Jiang XR, et al. Telomerase expression in human somatic cells does not induce changes associated with a transformed phenotype. Nat. Genet. 21: 111-114, 1999. PubMed: 9916802

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.
  • License agreement required for commercial customer uses.
  • This material is distributed for research purposes only. A signed addendum to the ATCC Material Transfer Agreement must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
FAQ's
  1. Number of population doublings (PDL) for ATCC® CRL-4001


    Date Updated: 3/27/2014

Restrictions

This material is subject to claims under U.S. Patent Nos. 6,261,836 and 6,337,200, other pending patent applications, and foreign counterparts thereof. It is required that either the Addendum for Commercial and For-Profit Organizations or the Addendum for Noncommercial and Academic Organizations, as appropriate, be signed and returned to ATCC before shipment.

References

Bodnar AG, et al. Extension of life-span by introduction of telomerase into normal human cells. Science 279: 349-352, 1998. PubMed: 9454332

Jiang XR, et al. Telomerase expression in human somatic cells does not induce changes associated with a transformed phenotype. Nat. Genet. 21: 111-114, 1999. PubMed: 9916802