| 產品名稱 |
hFOB 1.19 |
| 商品貨號 |
B164604 |
| 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]
Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country. |
| 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. |
| Disclosure |
This material is cited in a US or other Patent and may not be used to
infringe the claims. Depending on the wishes of the Depositor, ATCC may be
required to inform the Patent Depositor of the party to which the material was
furnished. This material may not have been produced or characterized by ATCC. |
| 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.
- Remove and discard culture medium.
-
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.
-
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.
-
Add 6.0 to 8.0 mL of complete growth medium and aspirate cells by gently pipetting.
-
Add appropriate aliquots of the cell suspension to new culture vessels.
- 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 |
| Temperature Effects |
Permissive temperature: 33.5°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 |
| Deposited As |
human |
| U.S. Patent Number |
|
| 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
|
