J Korean Orthop Res Soc.  1999 Oct;2(2):132-138.

Expression of Osteopontin in Osteoclast

• Chang JS
• Park JH
• Park YG
• Kim JH

Abstract

PURPOSE: This study was aimed to determine the relationship between osteopontin(OPN) and osteoclast, especially focused on whether ostecolast could produce osteopontin or not.
MATERIALS AND METHODS
Osteoclasts were isolated from the giant cell tumor of proximal tibia and seeded on the 13 mm round cover slip resided in 24 multi-well plates for culture. After 2 days, osteclasts on the cover slip were fixed with cold acetone for 3 minutes and immunocytochemistry was done with rabbit osteopontin antibody. For in situ RT-PCR, osteoclasts on the cover-slips were fixed with 4% paraformaldehyde for 4 hours and were treated to pepsin. PR-PCR was done and the PCR producst were stained with anti-digoxigenin-AP.
RESULTS
Osteopontins were found on the surface of the osteoclast by immunocytochemistry, and intense osteopontin mRNAs were found by in situ RT-PCR.
CONCLUSION
We have identified that osteoclast could synthesize the osteopontin, and confirmed that in situ RT-PCR was a very useful method in expressing small amount of mRNA in case of mixed cell culture. Further study was needed to identify the action of the osteopontin produced by the osteoclast.

MeSH Terms

Acetone
Cell Culture Techniques
Giant Cell Tumors
Immunohistochemistry
Osteoclasts*
Osteopontin*
Pepsin A
Polymerase Chain Reaction
RNA, Messenger
Tibia
Acetone
Osteopontin
Pepsin A
RNA, Messenger