Evans C E, Ng K, Allen J, Gallimore P
Department of Orthopaedic Surgery, University of Manchester.
J Orthop Res. 1995 May;13(3):317-24. doi: 10.1002/jor.1100130304.
At present, the majority of in vitro research into bone metabolism is performed on either primary cultures of bone or osteosarcoma lines. A better model of the behaviour of normal bone cells would be a cell line derived from normal, adult bone that retained osteoblast-like characteristics. We infected a culture of bone cells from adult humans with simian virus 1613, a variant of the simian virus 40, and obtained 12 clones of variable morphology. The clones were maintained in culture for as long as 6 months. Population doubling times, synthesis of alkaline phosphatase and osteocalcin, secretion of mineral, morphology, and ability to withstand freezing were examined. SV/EC cell morphology varied from the polygonal, osteoblast-like to the bipolar, fibroblast-like. Population doubling times ranged from 0.55 to 2.8 days (compared with 3.9 days for the nontransformed human osteoblast-like cells). Synthesis of alkaline phosphatase varied but was less than that by the human osteoblast-like cells. With the exception of clone 11, all of the transformed clones synthesised mineral in vitro under mineralising conditions. Four clones showed increased synthesis of alkaline phosphatase and increased population doubling times after passaging. All of the clones were successfully frozen and thawed, but, unlike normal human osteoblast-like cells, none responded to stimulation with parathyroid hormone.(ABSTRACT TRUNCATED AT 250 WORDS)
目前，大多数关于骨代谢的体外研究是在原代骨培养物或骨肉瘤细胞系上进行的。更能体现正常骨细胞行为的模型应该是源自正常成年骨且保留成骨细胞样特征的细胞系。我们用猿猴病毒40的一个变种——猿猴病毒1613感染了来自成年人类的骨细胞培养物，获得了12个形态各异的克隆。这些克隆在培养中维持了长达6个月。检测了群体倍增时间、碱性磷酸酶和骨钙素的合成、矿物质分泌、形态以及耐冻能力。SV/EC细胞形态从多边形、成骨细胞样到双极、成纤维细胞样不等。群体倍增时间在0.55至2.8天之间（未转化的人成骨细胞样细胞为3.9天）。碱性磷酸酶的合成各不相同，但低于人成骨细胞样细胞。除克隆11外，所有转化克隆在矿化条件下均能在体外合成矿物质。4个克隆在传代后碱性磷酸酶合成增加且群体倍增时间延长。所有克隆均成功冻存和解冻，但与正常的人成骨细胞样细胞不同，它们对甲状旁腺激素刺激均无反应。（摘要截短于250词）
