skip to main content
OSTI.GOV title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: Retrovirus-mediated conditional immortalization and analysis of established cell lines of osteoclast precursor cells

Abstract

Osteoclast precursor cells (OPCs) have previously been established from bone marrow cells of SV40 temperature-sensitive T antigen-expressing transgenic mice. Here, we use retrovirus-mediated gene transfer to conditionally immortalize OPCs by expressing temperature-sensitive large T antigen (tsLT) from wild type bone marrow cells. The immortalized OPCs proliferated at the permissive temperature of 33.5 deg. C, but stopped growing at the non-permissive temperature of 39 deg. C. In the presence of receptor activator of NF{kappa}B ligand (RANKL), the OPCs differentiated into tartrate-resistant acid phosphatase (TRAP)-positive cells and formed multinucleate osteoclasts at 33.5 deg. C. From these OPCs, we cloned two types of cell lines. Both differentiated into TRAP-positive cells, but one formed multinucleate osteoclasts while the other remained unfused in the presence of RANKL. These results indicate that the established cell lines are useful for analyzing mechanisms of differentiation, particularly multinucleate osteoclast formation. Retrovirus-mediated conditional immortalization should be a useful method to immortalize OPCs from primary bone marrow cells.

Authors:
 [1];  [1];  [2];  [1];  [1];  [1];  [1];  [3];  [4]
  1. Laboratory of Molecular Oncology, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101 (Japan)
  2. (Japan)
  3. Department of Neurodegenerative Disease, University College London, Square, London, WC1N 3BG (United Kingdom)
  4. Laboratory of Molecular Oncology, Nara Institute of Science and Technology, 8916-5 Takayama, Ikoma, Nara 630-0101 (Japan). E-mail: shishid@bs.naist.jp
Publication Date:
OSTI Identifier:
20854561
Resource Type:
Journal Article
Resource Relation:
Journal Name: Biochemical and Biophysical Research Communications; Journal Volume: 350; Journal Issue: 1; Other Information: DOI: 10.1016/j.bbrc.2006.09.020; PII: S0006-291X(06)02020-1; Copyright (c) 2006 Elsevier Science B.V., Amsterdam, The Netherlands, All rights reserved; Country of input: International Atomic Energy Agency (IAEA)
Country of Publication:
United States
Language:
English
Subject:
60 APPLIED LIFE SCIENCES; ACID PHOSPHATASE; ANTIGENS; BONE MARROW CELLS; GENES; LIGANDS; RECEPTORS; TRANSGENIC MICE

Citation Formats

Kawata, Shigehisa, Suzuki, Jun, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka 565-0871, Maruoka, Masahiro, Mizutamari, Megumi, Ishida-Kitagawa, Norihiro, Yogo, Keiichiro, Jat, Parmjit S., and Shishido, Tomoyuki. Retrovirus-mediated conditional immortalization and analysis of established cell lines of osteoclast precursor cells. United States: N. p., 2006. Web. doi:10.1016/j.bbrc.2006.09.020.
Kawata, Shigehisa, Suzuki, Jun, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka 565-0871, Maruoka, Masahiro, Mizutamari, Megumi, Ishida-Kitagawa, Norihiro, Yogo, Keiichiro, Jat, Parmjit S., & Shishido, Tomoyuki. Retrovirus-mediated conditional immortalization and analysis of established cell lines of osteoclast precursor cells. United States. doi:10.1016/j.bbrc.2006.09.020.
Kawata, Shigehisa, Suzuki, Jun, Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka 565-0871, Maruoka, Masahiro, Mizutamari, Megumi, Ishida-Kitagawa, Norihiro, Yogo, Keiichiro, Jat, Parmjit S., and Shishido, Tomoyuki. Fri . "Retrovirus-mediated conditional immortalization and analysis of established cell lines of osteoclast precursor cells". United States. doi:10.1016/j.bbrc.2006.09.020.
@article{osti_20854561,
title = {Retrovirus-mediated conditional immortalization and analysis of established cell lines of osteoclast precursor cells},
author = {Kawata, Shigehisa and Suzuki, Jun and Osaka University Graduate School of Medicine, 2-2 Yamada-oka, Suita, Osaka 565-0871 and Maruoka, Masahiro and Mizutamari, Megumi and Ishida-Kitagawa, Norihiro and Yogo, Keiichiro and Jat, Parmjit S. and Shishido, Tomoyuki},
abstractNote = {Osteoclast precursor cells (OPCs) have previously been established from bone marrow cells of SV40 temperature-sensitive T antigen-expressing transgenic mice. Here, we use retrovirus-mediated gene transfer to conditionally immortalize OPCs by expressing temperature-sensitive large T antigen (tsLT) from wild type bone marrow cells. The immortalized OPCs proliferated at the permissive temperature of 33.5 deg. C, but stopped growing at the non-permissive temperature of 39 deg. C. In the presence of receptor activator of NF{kappa}B ligand (RANKL), the OPCs differentiated into tartrate-resistant acid phosphatase (TRAP)-positive cells and formed multinucleate osteoclasts at 33.5 deg. C. From these OPCs, we cloned two types of cell lines. Both differentiated into TRAP-positive cells, but one formed multinucleate osteoclasts while the other remained unfused in the presence of RANKL. These results indicate that the established cell lines are useful for analyzing mechanisms of differentiation, particularly multinucleate osteoclast formation. Retrovirus-mediated conditional immortalization should be a useful method to immortalize OPCs from primary bone marrow cells.},
doi = {10.1016/j.bbrc.2006.09.020},
journal = {Biochemical and Biophysical Research Communications},
number = 1,
volume = 350,
place = {United States},
year = {Fri Nov 10 00:00:00 EST 2006},
month = {Fri Nov 10 00:00:00 EST 2006}
}