Loomer P M, Ellen R P, Tenenbaum H C
School of Dentistry, Department of Stomatology, Division of Periodontology, University of California, San Francisco, 521 Parnassus Avenue, Room C-628, San Francisco, CA 94143-0650, USA.
Cell Tissue Res. 1998 Oct;294(1):99-108. doi: 10.1007/s004410051160.
The processes involved in the regulation of bone cell metabolism are complex, including those implicated in bone cell coupling. This study was undertaken to develop a model that would permit real-time interaction between osteoclastic cells and osteoblasts in vitro. Osteogenic bone marrow stromal cells were isolated from 18-day-old embryonic chickens, while osteoclastic cells were isolated from laying White Leghorn hens on calcium-deficient diets. Osteoclastic cells (5x10(5)) were seeded onto mineral thin films and suspended above osteogenic cells (1x10(4)) already plated on the bottoms of tissue culture plate wells. The data showed that after 4 days of incubation there was up to a fivefold (P<0.05) reduction in all measured parameters of osteogenesis (mineralization, alkaline phosphatase activity and type I collagen production) in osteogenic cultures grown in the presence of osteoclastic cells. Similarly, osteoclastic cell-induced mineral resorption was reduced up to threefold (P<0.05). Co-culture effects on cellular responses could be manipulated by known antiresorptive agents (e.g., pamidronate) altering either the source or the age of osteoclastic cells. The results indicate that the co-culture model may be useful in the study of bone cell interactions.
骨细胞代谢调节过程复杂,包括与骨细胞偶联相关的过程。本研究旨在建立一个能够使破骨细胞与成骨细胞在体外实时相互作用的模型。从18日龄胚胎鸡中分离出成骨骨髓基质细胞,而破骨细胞则从缺钙饮食的产蛋白来航母鸡中分离。将破骨细胞(5×10⁵)接种到矿物薄膜上,并悬浮在已接种于组织培养板孔底部的成骨细胞(1×10⁴)上方。数据显示,培养4天后,在破骨细胞存在下生长的成骨培养物中,所有测量的成骨参数(矿化、碱性磷酸酶活性和I型胶原蛋白产生)最多降低了五倍(P<0.05)。同样,破骨细胞诱导的矿物质吸收最多降低了三倍(P<0.05)。已知的抗吸收剂(如帕米膦酸盐)可通过改变破骨细胞的来源或年龄来操纵共培养对细胞反应的影响。结果表明,共培养模型可能有助于骨细胞相互作用的研究。