hFOB 1.19細胞
運輸方式： 凍存運輸
細胞類型： 其他細胞類型
是否是腫瘤細胞： 0
物種來源： 人
數量： 大量
器官來源： 骨
生長狀態： 貼壁生長
年限： fetus
ATCC Number： CRL-11372?
Designations: hFOB 1.19
Depositors: SA Harris, TC Spelsberg
Biosafety Level: 2 [Cells contain SV40 viral sequences ]
Shipped: frozen
hFOB 1.19細胞Medium & Serum: See Propagation
Growth Properties: adherent
Organism: Homo sapiens deposited as human
Morphology:
Source: Organ: bone
Cell Type: osteoblast; SV40 large T antigen transfected
Cellular Products: alkaline phosphatase
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Antigen Expression: SV40 T antigen
DNA Profile (STR): 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
Cytogenetic Analysis: diploid, 43%; tetraploid, 57%
hFOB 1.19細胞Age: fetus
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).
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.
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.
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.
Propagation: ATCC 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). hFOB 1.19細胞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%.
Temperature: 34.0°C
Subculturing: Protocol:
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
Preservation: culture medium, 72%; additional fetal bovine serum, 20%; DMSO, 8%
Doubling Time: hFOB 1.19細胞36 hours at permissive temp of 33.5C
Related Products: recommended serum:ATCC 30-2020
References: 45062: Harris SA, Spelsberg TC. Immortalized human fetal osteoblastic cells. US Patent 5,681,701 dated Oct 28 1997
45223: 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
46124: Jacobs CR, et al. Differential effect of steady versus oscillating flow on bone cells. J. Biomech. 31: 969-976, 1998. PubMed: 9880053
