BJ (ATCC® CRL-2522)

Organism: Homo sapiens, human  /  Cell Type: fibroblast  /  Tissue: skin; foreskin  /  Disease: normal

Organism Homo sapiens, human
Tissue skin; foreskin
Cell Type fibroblast
Product Format frozen
Morphology fibroblast
Culture Properties adherent
Biosafety Level 1
Disease normal
Age newborn
Gender male
Applications
The cells may be used for stable transfection studies.
Storage Conditions liquid nitrogen vapor phase
Derivation
The line was established from skin taken from normal foreskin.
Clinical Data
newborn
male
Comments
The BJ cell line has a long lifespan in comparison with other normal human fibroblast cell lines.
A frozen ampule at population doubling 2.3 was received at the ATCC in April, 2000.
These cells have a reported normal diploid karyotype at population doubling 61 but an abnormal karyotype at population doubling 82.
They are telomerase negative.
Cells from ATCC ampules have the capacity to proliferate to a maximum of 72 population doublings before the onset of senescence.
Complete Growth Medium The base medium for this cell line is ATCC-formulated Eagle's Minimum Essential Medium, Catalog No. 30-2003. To make the complete growth medium, add the following components to the base medium: fetal bovine serum to a final concentration of 10%.
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 0.25% (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 1:2 to 1:9 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°C
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 Capacity Cells from ATCC ampules have the capacity to proliferate to a maximum of 72 population doublings before the onset of senescence.
Name of Depositor JR Smith
Passage History
A frozen ampule at population doubling 2.3 was received at the ATCC in April, 2000.
References

Yi X, et al. Both transcriptional and posttranscriptional mechanisms regulate human telomerase template RNA levels. Mol. Cell. Biol. 19(6): 3989-3997, 1999. PubMed: 10330139

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

Huffman KE, et al. Telomere shortening is proportional to the size of the G-rich telomeric 3'-overhang. J. Biol. Chem. 275: 19719-19722, 2000. PubMed: 10787419

Morales CP, et al. Absence of cancer-associated changes in human fibroblasts immortalized with telomerase. Nat. Genet. 21: 115-118, 1999. PubMed: 9916803

Basic Documentation
References

Yi X, et al. Both transcriptional and posttranscriptional mechanisms regulate human telomerase template RNA levels. Mol. Cell. Biol. 19(6): 3989-3997, 1999. PubMed: 10330139

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

Huffman KE, et al. Telomere shortening is proportional to the size of the G-rich telomeric 3'-overhang. J. Biol. Chem. 275: 19719-19722, 2000. PubMed: 10787419

Morales CP, et al. Absence of cancer-associated changes in human fibroblasts immortalized with telomerase. Nat. Genet. 21: 115-118, 1999. PubMed: 9916803