Detection of osteoclastic cell-cell fusion through retroviral vector packaging

Takako Kondo, Kyoji Ikeda, Koichi Matsuo

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Cell-cell fusion generates multinucleated cells such as osteoclasts in bone, myotubes in muscle, and trophoblasts in placenta. Molecular details governing these fusion processes are still largely unknown. As a step toward identification of fusogenic genes, we tested the concept that retroviral vectors can be packaged as a result of cell-cell fusion. First, we introduced replication-deficient retroviral vectors expressing mCAT-1, which mediates fusogenic interaction with the retroviral envelope protein Env, into Chinese hamster ovary (CHO) cells to generate vector cells. Plasmids expressing virion proteins Gag, Pol, and Env were introduced into a separate culture of CHO cells to generate packaging cells. Co-culturing vector and packaging cells resulted in production of infectious retroviruses carrying the mCAT-1 gene as a consequence of cell-cell fusion. Second, we introduced a retroviral vector into primary osteoclast precursors and co-cultured them with established osteoclast precursor RAW264.7 cells, which turned out to harbor packaging activity. Packaged retroviral vector was detected in culture supernatants only where the osteoclast differentiation factor receptor activator for NF-κB ligand (RANKL) induced fusion between these two cell types. These data suggest that retrovirus production can occur as a result of cell-cell fusion. This provides a novel approach for isolating and characterizing fusogenic genes using retroviral expression vectors.

Original languageEnglish
Pages (from-to)1120-1126
Number of pages7
JournalBone
Volume35
Issue number5
DOIs
Publication statusPublished - 2004 Nov

Fingerprint

Cell Fusion
Product Packaging
Osteoclasts
Retroviridae
Cricetulus
Ovary
gag-pol Fusion Proteins
env Gene Products
RANK Ligand
Genes
Skeletal Muscle Fibers
Trophoblasts
Virion
Placenta
Plasmids

Keywords

  • Fusion
  • Osteoclasts
  • Retrovirus

ASJC Scopus subject areas

  • Physiology
  • Hematology

Cite this

Detection of osteoclastic cell-cell fusion through retroviral vector packaging. / Kondo, Takako; Ikeda, Kyoji; Matsuo, Koichi.

In: Bone, Vol. 35, No. 5, 11.2004, p. 1120-1126.

Research output: Contribution to journalArticle

Kondo, Takako ; Ikeda, Kyoji ; Matsuo, Koichi. / Detection of osteoclastic cell-cell fusion through retroviral vector packaging. In: Bone. 2004 ; Vol. 35, No. 5. pp. 1120-1126.
@article{dc816d300e2c48e1ac3dce63eb4bf5f1,
title = "Detection of osteoclastic cell-cell fusion through retroviral vector packaging",
abstract = "Cell-cell fusion generates multinucleated cells such as osteoclasts in bone, myotubes in muscle, and trophoblasts in placenta. Molecular details governing these fusion processes are still largely unknown. As a step toward identification of fusogenic genes, we tested the concept that retroviral vectors can be packaged as a result of cell-cell fusion. First, we introduced replication-deficient retroviral vectors expressing mCAT-1, which mediates fusogenic interaction with the retroviral envelope protein Env, into Chinese hamster ovary (CHO) cells to generate vector cells. Plasmids expressing virion proteins Gag, Pol, and Env were introduced into a separate culture of CHO cells to generate packaging cells. Co-culturing vector and packaging cells resulted in production of infectious retroviruses carrying the mCAT-1 gene as a consequence of cell-cell fusion. Second, we introduced a retroviral vector into primary osteoclast precursors and co-cultured them with established osteoclast precursor RAW264.7 cells, which turned out to harbor packaging activity. Packaged retroviral vector was detected in culture supernatants only where the osteoclast differentiation factor receptor activator for NF-κB ligand (RANKL) induced fusion between these two cell types. These data suggest that retrovirus production can occur as a result of cell-cell fusion. This provides a novel approach for isolating and characterizing fusogenic genes using retroviral expression vectors.",
keywords = "Fusion, Osteoclasts, Retrovirus",
author = "Takako Kondo and Kyoji Ikeda and Koichi Matsuo",
year = "2004",
month = "11",
doi = "10.1016/j.bone.2004.06.011",
language = "English",
volume = "35",
pages = "1120--1126",
journal = "Bone",
issn = "8756-3282",
publisher = "Elsevier Inc.",
number = "5",

}

TY - JOUR

T1 - Detection of osteoclastic cell-cell fusion through retroviral vector packaging

AU - Kondo, Takako

AU - Ikeda, Kyoji

AU - Matsuo, Koichi

PY - 2004/11

Y1 - 2004/11

N2 - Cell-cell fusion generates multinucleated cells such as osteoclasts in bone, myotubes in muscle, and trophoblasts in placenta. Molecular details governing these fusion processes are still largely unknown. As a step toward identification of fusogenic genes, we tested the concept that retroviral vectors can be packaged as a result of cell-cell fusion. First, we introduced replication-deficient retroviral vectors expressing mCAT-1, which mediates fusogenic interaction with the retroviral envelope protein Env, into Chinese hamster ovary (CHO) cells to generate vector cells. Plasmids expressing virion proteins Gag, Pol, and Env were introduced into a separate culture of CHO cells to generate packaging cells. Co-culturing vector and packaging cells resulted in production of infectious retroviruses carrying the mCAT-1 gene as a consequence of cell-cell fusion. Second, we introduced a retroviral vector into primary osteoclast precursors and co-cultured them with established osteoclast precursor RAW264.7 cells, which turned out to harbor packaging activity. Packaged retroviral vector was detected in culture supernatants only where the osteoclast differentiation factor receptor activator for NF-κB ligand (RANKL) induced fusion between these two cell types. These data suggest that retrovirus production can occur as a result of cell-cell fusion. This provides a novel approach for isolating and characterizing fusogenic genes using retroviral expression vectors.

AB - Cell-cell fusion generates multinucleated cells such as osteoclasts in bone, myotubes in muscle, and trophoblasts in placenta. Molecular details governing these fusion processes are still largely unknown. As a step toward identification of fusogenic genes, we tested the concept that retroviral vectors can be packaged as a result of cell-cell fusion. First, we introduced replication-deficient retroviral vectors expressing mCAT-1, which mediates fusogenic interaction with the retroviral envelope protein Env, into Chinese hamster ovary (CHO) cells to generate vector cells. Plasmids expressing virion proteins Gag, Pol, and Env were introduced into a separate culture of CHO cells to generate packaging cells. Co-culturing vector and packaging cells resulted in production of infectious retroviruses carrying the mCAT-1 gene as a consequence of cell-cell fusion. Second, we introduced a retroviral vector into primary osteoclast precursors and co-cultured them with established osteoclast precursor RAW264.7 cells, which turned out to harbor packaging activity. Packaged retroviral vector was detected in culture supernatants only where the osteoclast differentiation factor receptor activator for NF-κB ligand (RANKL) induced fusion between these two cell types. These data suggest that retrovirus production can occur as a result of cell-cell fusion. This provides a novel approach for isolating and characterizing fusogenic genes using retroviral expression vectors.

KW - Fusion

KW - Osteoclasts

KW - Retrovirus

UR - http://www.scopus.com/inward/record.url?scp=8444227794&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=8444227794&partnerID=8YFLogxK

U2 - 10.1016/j.bone.2004.06.011

DO - 10.1016/j.bone.2004.06.011

M3 - Article

C2 - 15542037

AN - SCOPUS:8444227794

VL - 35

SP - 1120

EP - 1126

JO - Bone

JF - Bone

SN - 8756-3282

IS - 5

ER -