저칼슘식이로 사육한 생쥐의 골수세포 배양에 의한 파골세포양세포의 형성
Osteoclast-Like Cell Formation in Vitro from Calcium-Deficient Mouse Bone Marrow Cells
소속 상세정보
박주철
강선주/임도선/김현만/고재승
KMID : 0355819950190010107
Abstract
Although it is now well established that osteoclasts are derived from mononuclear hematopoietic stem cells, most likely within the granulocyte-macrophage lineage, the exact differentiation process of osteolast precusors has only been partially
discovered. This study was carried out to investigate the following topics by examining the osteoclast-like cell formation from the calcium-deficient mouse bone marrow cells; 1) the effect of calcium deficiency on the formation of the mononuclear
phagocytes and the mononuclear precusors of the osteoclast in the mouse bone marrow; 2) the effect of TNF (tumor necrosis factor)-a on the differentiation of osteoclast-like cells from the calcium-deficient mouse bone marrow culture; 3) the
formation of
the osteoclast-like cell from the calcium-deficient mouse bone marrow mononuclear cells and the relationship between the osteoblast cell line(MC3T3E1) and osteoclast-like cell formation of the osteoclast-like cells from the mononuclear cells
which
were
collected from PTH-pretreated mouse bone marrow cells to investigate the relationship between PTH and calcium deficiency in osteoclast-like cell formation.
Bone marrow cells were isolated from marrow of mouse feeding normal diet or calcium deficient diet for 7 and 14 days. Bone marrow mononuclear cells were fractionated by cetrifugation on Hypaque-Ficoll density gradients. PTH-pretreated mouse bone
marrow
mononuclear cells were obtained by collecting the nonadherent cells after the culture of normal mouse marrow cells with PTH for 6 days.
The prepared cells were cultured on dentin disc at the concentration of 1.5-2 X10E6 cells per 0.5ml of a-MEM containing 10% FCS for 3, 5 and days. Thereafter, TRAP staining, NSE staining, effect of calcitonin on the osteoclast-like cell and
scanning
electron microscope analysis of resorption lacunae on the dentin disc were performed
@ES The results were as follows ;
@EN 1. Number of the osteoclast precusors and mononuclear phagocytes in bone marrow was increased by calciumdeficient diet.
2. more osteoclast-like cells were formed from the calcium-deficient mouse bone marrow cells than from normal cells.
3. Formation of osteoclast-like cells was significantly stimulated by TNF-a in normal mouse bone marrow cells and calcium-deficient mouse bone marrow cells at concentration of 10ng/ml and 100ng/ml.
4. Osteoblast cell line(MC3T3E1) did not play an important role in osteoclast-like cell formation.
5. Formation of the osteoclast-like cells from the mononuclear cells which were collected from PTH-pretreated mouse bone marrow cells was similar to that of calcium-deficient mouse bone marrow mononuclear cells, which suggests that the effect of
calcium deficiency in osteoclast formation may be mediated by PTH.
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