DOI: 10.1002/jbmr.5650100203. Development and characterization of a conditionally immortalized human fetal osteoblastic cell line

Development and characterization of a conditionally immortalized human fetal osteoblastic cell line

10.1002/jbmr.5650100203

Crossref journal-article

Oxford University Press (OUP)

Journal of Bone and Mineral Research (286)

Abstract

Abstract We report the establishment of a human fetal osteoblast cell line derived from biopsies obtained from a spontaneous miscarriage. Primary cultures isolated from fetal tissue were transfected with a gene coding for a temperature-sensitive mutant (tsA58) of SV40 large T antigen along with a gene coding for neomycin (G418) resistance. Individual neomycin resistant colonies were screened for alkaline phosphatase (AP)-specific staining. The clone with the highest AP level, hFOB 1.19, was examined further for other osteoblast phenotypic markers. Incubation of hFOB cells at the permissive temperature (33.5°C) resulted in rapid cell division, whereas little or no cell division occurred at the restrictive temperature (39.5°C). Both AP activity and osteocalcin (OC) secretion increased in a dose-dependent manner following dihydroxyvitamin D3 (1,25-D3) treatment when cultured at either temperature. However, AP and 1,25-D3-induced OC levels were elevated in confluent hFOB cells cultured at 39.5°C compared with 33.5°C. Treatment of hFOB cells with 1–34 parathyroid hormone (PTH) resulted in an increase in cAMP levels. Upon reaching confluence, hFOB cultures went through programmed differentiation and formed mineralized nodules as observed by von Kossa staining. Further, immunostaining of postconfluent, differentiated hFOB cells showed that high levels of osteopontin, osteonectin, bone sialoprotein, and type I collagen were expressed. Therefore, the clonal cell line hFOB 1.19 provides a homogeneous, rapidly proliferating model system to study certain stages of human osteoblast differentiation.

Bibliography

Dr. Harris, S. A., Enger, R. J., Riggs, L. B., & Spelsberg, T. C. (1995). Development and characterization of a conditionally immortalized human fetal osteoblastic cell line. Journal of Bone and Mineral Research, 10(2), 178â186.

Authors 4

Steven A. Dr. Harris (first)
Robert J. Enger (additional)
Lawrence B. Riggs (additional)
Thomas C. Spelsberg (additional)

References 23 Referenced 371

10.1016/0221-8747(84)90064-X / Metab Bone Dis Res / Production of osteocalcin by human bone cells in vitro. Effects of 1,25(OH)2D3, 24,25(OH)D3, PTH, and glucocorticoids by Beresford (1984)
10.1007/BF02557826 / Calcif Tissue Int / Human bone cells in vitro by Robey (1985)
10.1210/endo-100-5-1357 / Endocrinology / Evidence for preferential effects of parathyroid hormone, calcitonin and adenosine on bone and periosteum by Peck (1977)
10.1002/jcp.1041230316 / J Cell Physiol / 1α, 25-dihy-droxyvitamin D3 specific regulation of growth, morphology, and fibronectin in a human osteosarcoma cell line by Franceschi (1985)
10.1038/319511a0 / Nature / A human osteosarcoma cell line secretes a growth factor structurally related to a homodimer of PDGF A-chains by Heldin (1986)
{'key': '2024013114222426900_bib6', 'first-page': '4961', 'article-title': 'Characterization of a human cell line (Saos-2) with osteoblastic properties', 'volume': '47', 'author': 'Rodan', 'year': '1987', 'journal-title': 'Cancer Res'} / Cancer Res / Characterization of a human cell line (Saos-2) with osteoblastic properties by Rodan (1987)
10.1002/jcp.1041340321 / J Cell Physiol / Tumor necrosis factor stimulates proliferation of human osteosarcoma cells and accumulation of c-myc messenger RNA by Kirstein (1988)
{'key': '2024013114222426900_bib8', 'first-page': '244', 'volume-title': 'Bone and Mineral Research', 'author': 'Rodan', 'year': '1984'} / Bone and Mineral Research by Rodan (1984)
10.1016/0076-6879(87)45019-2 / Meth Enzymol / Hormonal influences on bone cells by Martin (1987)
{'key': '2024013114222426900_bib10', 'first-page': '85', 'article-title': 'Gene expression in osteoblastic cells', 'volume': '1', 'author': 'Rodan', 'year': '1991', 'journal-title': 'Crit Rev Eukaryot Gene Expr'} / Crit Rev Eukaryot Gene Expr / Gene expression in osteoblastic cells by Rodan (1991)
10.1002/jbmr.5650070203 / J Bone Miner Res / Development and characterization of a rapidly proliferating, well-differentiated cell line derived from normal adult human osteoblast-like cells trans-fected with SV40 large T antigen by Keeting (1992)
10.1111/j.1349-7006.1993.tb02869.x / Jpn J Cancer Res / Establishment and characterization of a simian virus 40-immortalized osteoblastic cell line from normal human bone by Chiba (1993)
10.1128/jvi.15.3.613-618.1975 / J Virol / Function of simian virus 40 gene A transforming infection by Tegtmeyer (1975)
10.1016/0092-8674(88)90559-4 / Cell / SV40 large tumor antigen forms a specific complex with the product of the retinoblastoma susceptibility gene by DeCaprio (1988)
10.1016/0092-8674(89)90983-5 / Cell / SV40 large T antigen binds preferentially to an under-phosphorylated member of the retinoblastoma susceptibility gene product family by Ludlow (1989)
10.1210/mend-3-10-1511 / Mol Endocrinol / Differentiated mammalian cell lines immortalized by temperature-sensitive tumor viruses by Chou (1989)
10.1002/jcp.1041430203 / J Cell Physiol / Factors that promote progressive development of the osteoblast phenotype in cultured fetal rat calvaria cells by Aronow (1989)
10.1096/fasebj.4.13.2210157 / FASEB J / Relationship of cell growth to the regulation of tissue-specific gene expression during osteoblast differentiation by Stein (1990)
10.1016/S0021-9258(18)47991-4 / J Biol Chem / Purification and partial characterization of small proteoglycans I and II, bone sialoproteins I and II, and osteonectin from the mineral compartment of developing human bone by Fisher (1987)
{'key': '2024013114222426900_bib20', 'first-page': '5365', 'article-title': 'Conditional immortalization of human thyroid epithelial cells: A tool for analysis of oncogene action', 'volume': '10', 'author': 'Wynford-Thomas', 'year': '1990', 'journal-title': 'Mol Cell Biol'} / Mol Cell Biol / Conditional immortalization of human thyroid epithelial cells: A tool for analysis of oncogene action by Wynford-Thomas (1990)
{'key': '2024013114222426900_bib21', 'first-page': '327', 'article-title': 'Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter', 'volume': '1', 'author': 'Southern', 'year': '1982', 'journal-title': 'J Mol Appl Gen'} / J Mol Appl Gen / Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter by Southern (1982)
10.1016/0165-4608(88)90312-3 / Cancer Genet Cytogenet / Culturing and robotic harvesting of bone marrow, lymph nodes, peripheral blood, fibroblasts, and solid tumors with in situ techniques by Spurbeck (1988)
{'key': '2024013114222426900_bib23', 'first-page': '1', 'volume-title': 'Methods of Calcified Tissue Preparation', 'author': 'Schenk', 'year': '1984'} / Methods of Calcified Tissue Preparation by Schenk (1984)

Dates

Type	When
Created	15 years, 7 months ago (Feb. 3, 2010, 11:03 a.m.)
Deposited	1 year, 5 months ago (March 13, 2024, 10:59 a.m.)
Indexed	1 month ago (Aug. 6, 2025, 8:16 a.m.)
Issued	30 years, 7 months ago (Feb. 1, 1995)
Published	30 years, 7 months ago (Feb. 1, 1995)
Published Online	30 years, 7 months ago (Feb. 1, 1995)
Published Print	30 years, 7 months ago (Feb. 1, 1995)

Funders 1

Mayo Foundation and from NIH/NIAMS
Awards1
1. AR41652

BibTeX

@article{Dr_Harris_1995, title={Development and characterization of a conditionally immortalized human fetal osteoblastic cell line}, volume={10}, ISSN={1523-4681}, url={http://dx.doi.org/10.1002/jbmr.5650100203}, DOI={10.1002/jbmr.5650100203}, number={2}, journal={Journal of Bone and Mineral Research}, publisher={Oxford University Press (OUP)}, author={Dr. Harris, Steven A. and Enger, Robert J. and Riggs, Lawrence B. and Spelsberg, Thomas C.}, year={1995}, month=feb, pages={178–186} }

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