hFOB 1.19 (ATCC® CRL-11372)

Organism: Homo sapiens, human  /  Cell Type: osteoblast; SV40 large T antigen transfected  /  Tissue: bone  / 

Organism Homo sapiens, human
Tissue
bone
Cell Type osteoblast; SV40 large T antigen transfected
Product Format frozen
Culture Properties adherent
Biosafety Level 2 [Cells contain SV40 viral sequences]
Age fetus
Applications The cells provide a homogenous, rapidly proliferating model system for studying normal human osteoblast differentiation, osteoblast physiology, and hormonal, growth factor, and other cytokine effects on osteoblast function and differentiation.
Karyotype diploid, 43%; tetraploid, 57%
Derivation
This line was established by transfection of limb tissue obtained from a spontaneous miscarriage with the temperature sensitive expression vector pUCSVtsA58 and the neomycin resistance expression vector pSV2-neo. Clones were selected in the presence of 0.6 mg/mL G418.
Antigen Expression
SV40 T antigen
Genes Expressed
alkaline phosphatase
Cellular Products
alkaline phosphatase
Comments
Cells grown at a permissive temperature of 33.5°C exhibit rapid cell division (Doubling time of 36 hrs), whereas little cell division occurs at a restrictive temperature of 39.5°C (Doubling time of 96 hrs).
The cells have the ability to differentiate into mature osteoblasts expressing the normal osteoblast phenotype. At the restrictive temperatures, cell division is slowed, differentiation increases, and a more mature osteoblast phenotype is produced.
Complete Growth Medium The base medium for this cell line is a 1:1 mixture of Ham's F12 Medium Dulbecco's Modified Eagle's Medium,with 2.5 mM L-glutamine (without phenol red). To make the complete growth medium, add the following components to the base medium: 0.3 mg/ml G418; 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 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 34°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 34°C.
Subcultivation Ratio: A subcultivation ratio of 1:4 is recommended
Medium Renewal: Every 2 to 3 days
Cryopreservation
culture medium, 72%; additional fetal bovine serum, 20%; DMSO, 8%
Culture Conditions
Temperature: 34°C
STR Profile
Amelogenin: X
CSF1PO: 10,13
D13S317: 11,12
D16S539: 9,13
D5S818: 11,12
D7S820: 8,10
THO1: 7,9.3
TPOX: 11
vWA: 16,18
Population Doubling Time 36 hours at permissive temp of 33.5°C
Name of Depositor SA Harris, TC Spelsberg
References

Harris SA, Spelsberg TC. Immortalized human fetal osteoblastic cells. US Patent 5,681,701 dated Oct 28 1997

Harris SA, et al. Development and characterization of a conditionally immortalized human fetal osteoblastic cell line. J. Bone Miner. Res. 10: 178-186, 1995. PubMed: 7754797

Jacobs CR, et al. Differential effect of steady versus oscillating flow on bone cells. J. Biomech. 31: 969-976, 1998. PubMed: 9880053

Basic Documentation
References

Harris SA, Spelsberg TC. Immortalized human fetal osteoblastic cells. US Patent 5,681,701 dated Oct 28 1997

Harris SA, et al. Development and characterization of a conditionally immortalized human fetal osteoblastic cell line. J. Bone Miner. Res. 10: 178-186, 1995. PubMed: 7754797

Jacobs CR, et al. Differential effect of steady versus oscillating flow on bone cells. J. Biomech. 31: 969-976, 1998. PubMed: 9880053