Tumor Necrosis Factor-α와 저칼슘식이가 생쥐 골수세포의 파골세포양세포 분화에 미치는 영향
Tomor Necrosis Factor-α Stimulates Osteoclast-Like Cell Formation in the Calcium-Deficient Mouse Marrow Cell Culture
김영설, 강선주, 박귀원, 김석화,
소속 상세정보
김영설 ( ) - 서울대학교
강선주 ( ) - 서울대학교
박귀원 ( ) - 서울대학교
김석화 ( ) - 서울대학교
KMID : 0355819930170010075
Abstract
To study the effect of tumor necrosis factor-a and calcium deficiency on osteoclast generation from the precursor cells, the bone marrow cells were prepared from mice(ICR) fed a low calcium diet for 2 weeks /and 3 weeks, and normal diet. The
tibia
was
dissected aseptically and the marrow cavity was flushed with 1ml of a-minimum essential medium by slow injection. The collected marrow cells were cultured at 1.5x10E6 cells/well in 4 well plate for 3 days, 6 days, and 8 days. in experimental
group,
recombinant mouse TNF-a 100ng/ml was added. After cultures, staining for tartrate-resistant acid phosphatase(TRACT)-marker enzyme of osteoclast-were performed according to the modified method of Burstone. The TRACP-positive mononuclear cells and
multinucleated cells, which have 3 or more nuclei, were counted.
@ES The observed results were as follows.
@EN 1. Few TRACT-positive mononuclear cells and TRACP positive multinucleated cells appeared in the culture of bone marrow cells from mouse fed normal diet /and low calcium diet.
2. TNF-a stimulated the formation of TRACP-positive mononuclear cells and TRACT-positive multinucleated cells in the culture of bone marrow cells from mouse fed normal diet /and low calcium diet.
3. Low calcium diet potentiated the generation of TRACP-positive mononuclear cells and TRACP-positive multinucleated cells induced by TNF-a.
4. There types of TRACP-positive multinucleated cells were detected: one in which nuclei were located in the center of the cell, the other in which nuclei were located in the periphery of the cel, and the third in which nuclei were scattered.
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