Rat Calvarial Cell Lines Immortalized with SV-40 Large T Antigen: Constitutive and Retinoic Acid- Inducible Expression of Osteoblastic Features
- 1 June 1989
- journal article
- research article
- Published by The Endocrine Society in Endocrinology
- Vol. 124 (6) , 3060-3068
- https://doi.org/10.1210/endo-124-6-3060
Abstract
Two new bone cell lines were established by immortalizing cells derived from embryonic rat calvariae with a recombinant retrovirus containing the cDNA for SV-40 large T antigen and the neomycin resistance gene. One cell line, RCT-1, isolated from early digest cells, a population which typically does not express osteoblastic features, displayed osteobastic characteristics only after 3 days of treatment with 1 .mu.M retinoic acid: alkaline phosphatase activity increased from 0.003 to 0.25 .mu.mol/min .cntdot. mg protein, the steady state level of type I procollagen mRNA increased 4-fold, and the cells acquired a PTH-stimulable adenylate cyclase (EC50, 10 nM). mRNA for osteopontin, an abundant bone matrix protein, was induced in RCT-1 cells by 1,25-dihydroxyvitamin D3 (10 nM). The second cell line, RCT-3, isolated from late digest cells, a population previously shown to be enriched with differentiated osteoblasts, expressed constitutively the properties described above. In addition, RCT-3 cells responded to interleukin-1 by increased prostaglandin production (EC50, 20 pM) and to prostaglandin E2 by enhanced cAMP accumulation, features exhibited by calvarial cells in organ culture. Thus, the SV-40 immortalized cell lines we describe retained many of the characteristics of osteoblasts in primary culture, including hormonal regulation of phenotype-related genes. In RCT-1 cells the coordinate induction of several properties by retinoic acid offers a new model for the study of differentiation-related gene expression in bone cells.This publication has 37 references indexed in Scilit:
- In vitro differentiation and calcification in a new clonal osteogenic cell line derived from newborn mouse calvaria.The Journal of cell biology, 1983
- The effect of 1,25(OH)2D3 on alkaline phosphatase in osteoblastic osteosarcoma cells.Journal of Biological Chemistry, 1982
- Isolation of bone cell clones with differences in growth, hormone responses, and extracellular matrix production.The Journal of cell biology, 1982
- Rabbit cranial suture fibroblasts under tension express a different collagen phenotypeArchives of Oral Biology, 1981
- Osteonectin, a bone-specific protein linking mineral to collagenCell, 1981
- Regulation of terminal differentiation of cultured human keratinocytes by vitamin APublished by Elsevier ,1981
- 1,25-Dihydroxyvitamin D3 increases synthesis of the vitamin K-dependent bone protein by osteosarcoma cells.Journal of Biological Chemistry, 1980
- Refinement of the Coomassie blue method of protein quantitationAnalytical Biochemistry, 1978
- Evidence for Preferential Effects of Parathyroid Hormone, Calcitonin and Adenosine on Bone and Periosteum1Endocrinology, 1977
- The estimation of two collagens from human dermis by interrupted gel electrophoresisBiochemical and Biophysical Research Communications, 1976