RRC ID 12031
Author Kawata S, Suzuki J, Maruoka M, Mizutamari M, Ishida-Kitagawa N, Yogo K, Jat PS, Shishido T.
Title Retrovirus-mediated conditional immortalization and analysis of established cell lines of osteoclast precursor cells.
Journal Biochem Biophys Res Commun
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 degrees C, but stopped growing at the non-permissive temperature of 39 degrees C. In the presence of receptor activator of NFkappaB ligand (RANKL), the OPCs differentiated into tartrate-resistant acid phosphatase (TRAP)-positive cells and formed multinucleate osteoclasts at 33.5 degrees 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.
Volume 350(1)
Pages 97-104
Published 2006-11-10
DOI 10.1016/j.bbrc.2006.09.020
PII S0006-291X(06)02020-1
PMID 17005159
MeSH Animals Cell Differentiation* / drug effects Cell Survival Mice Osteoclasts / cytology* Osteoclasts / drug effects Osteoclasts / metabolism RANK Ligand / pharmacology Retroviridae / genetics* Stem Cells / cytology* Stem Cells / drug effects Temperature
IF 2.985
Times Cited 3
DNA material pCAhMCSF Human M-CSF(RDB01524)