Robey Pamela Gehron, Termine John D
Mineralized Tissue Research Branch, National Institute of Dental Research, National Institutes of Health, Bldg 30 Rm 214, 20205, Bethesda, Maryland, USA.
Calcif Tissue Int. 1985 Sep;37(5):453-460. doi: 10.1007/BF02557826.
Human bone cell cultures were established by maintaining collagenase-treated, bone fragments in low Ca medium. The resulting cell cultures exhibited a high level of alkaline phosphatase activity and produced a significant increase in intracellular cAMP when exposed to the 1-34 fragment of human parathyroid hormone. With continued culture, the cells formed a thick, extracellular matrix that mineralized when cultures were provided daily with normal levels of calcium, fresh ascorbic acid (50 μg/ml) and 10 mM β-glycerol phosphate. Biosynthetically, these cells produced type I collagen (without any type III collagen), and the bone-specific protein, osteonectin. In addition, the cells produced sulfated macromolecules electrophoretically identical to those positively identified as the bone proteoglycan in parallel cultures of fetal bovine bone cells. This technique provides a useful system for the study of osteoblast metabolismin vitro.
通过在低钙培养基中培养经胶原酶处理的骨碎片来建立人骨细胞培养物。所得细胞培养物表现出高水平的碱性磷酸酶活性，并且当暴露于人甲状旁腺激素的1-34片段时，细胞内cAMP显著增加。随着培养的继续，细胞形成了厚厚的细胞外基质，当每天为培养物提供正常水平的钙、新鲜抗坏血酸（50μg/ml）和10mMβ-甘油磷酸时，该基质会矿化。从生物合成角度来看，这些细胞产生I型胶原蛋白（无任何III型胶原蛋白）以及骨特异性蛋白骨连接蛋白。此外，这些细胞产生的硫酸化大分子在电泳上与在胎牛骨细胞平行培养物中被确认为骨蛋白聚糖的大分子相同。该技术为体外研究成骨细胞代谢提供了一个有用的系统。
