An In Vitro Study of Osteoclast Formation from Peripheral Blood Monocytes

周邊血液單核球形成破骨細胞的體外研究

史中(Chung Shih)

《Journal of Medical Sciences》 16卷6期 (1996/06) Pp. 389-400

摘要

有關破骨細胞的形成，由文獻得知骨髓細胞可在動物體內或體外形成破骨細胞，但周邊血液單核球是否可以在體外培養形成破骨細胞，一直未有文獻報導。本研究旨在證實周邊血液單核球在培養血中與骨組織共同培養可以形成破骨細胞。實驗方法：自懷孕13-14天的BALB/C 種小白鼠中取得不含破骨細胞的胎鼠頭蓋骨組織，然後使用BGJb培養被進行培養9天，再使用Ficoll-Paque比重梯度分離法獲得周邊血液單核球並加入實驗組培養血中與頭蓋骨組織共同培養的天。實驗結果，在加有單核細胞（單核球）的實驗組培養血中發現多核的破骨細胞形成並貼附在骨表面。另外，平均每10萬個周邊血液小比重單核細胞可形成1.4個破骨細胞，並且在對照組培養血中並沒有任何破骨細胞的出現。本研究結果顯示：（1）周邊血液單核球可在體外培養之骨組織表面聚合形成破骨細胞和（2）新建立的體外研究模式可應用來探討和評估破骨細胞形成和功能方面的研究。

關鍵字

頭蓋骨組織 ； 單核球 ； 破骨細胞 ； 刷狀緣

並列摘要

Previous studies have shown that osteoclasts are derived from monocytes of hemopoietic bone marrow both in vivo and in vitro and from peripheral blood only in vivo. The purpose of this study was to demonstrate that multinucleated osteoclasts could develop after adding monocytes from peripheral blood into established explants of fetal mouse calvaria in vitro. In order to ultilize osteoclast-free bone, the fetal calvaria was obtained from 13-14 day pregnant BALB/C mice and cultured in BGJb medium for 9 days. At day 9, peripheral blood mononuclear cells were isolated as a light density layer from adult BALB/C mice with the Ficoll-Paque density gradient separation method and co-cultured with the osteoclast-free, fetal mouse calvaria. After 10 days of co-culture, multinucleated cells which have all the characteristics of osteoclasts were found in juxtaposition to seams of bone. In addition, an average of 1.4 osteoclasts per 1 X 10^5 peripheral blood light density mononuclear cells were formed in the experimental group while none was found in the mononuclear cell-free control group. It is concluded that 1) peripheraI blood monocytes could give rise to multinucleated osteoclasts in developing bone in vitro and 2) a new and suitable model for in vitro study of osteoclast formation and function was established.

並列關鍵字

Calvaria ； monocyte ； osteoclast ； ruffled border