Expression of Osteopontin in Osteoclast.
- Author:
Jae Suk CHANG
;
Jong Hoon PARK
;
Yong Gu PARK
;
Jeong Hwa KIM
- Publication Type:Original Article
- MeSH:
Acetone;
Cell Culture Techniques;
Giant Cell Tumors;
Immunohistochemistry;
Osteoclasts*;
Osteopontin*;
Pepsin A;
Polymerase Chain Reaction;
RNA, Messenger;
Tibia
- From:Journal of Korean Orthopaedic Research Society
1999;2(2):132-138
- CountryRepublic of Korea
- Language:Korean
-
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.
