Osteocytes are of high importance in bone metabolism as they orchestrate bone remodeling, react to mechanosensory stimuli and have endocrine functions. In vitro investigations with osteocytes are therefore of high relevance for biomaterial and drug testing. The application of primary human cells instead of rodent osteocyte cell lines like MLOY4 and IDG SW3 is desirable but provides the challenge of isolating these cells, which are deeply embedded into the mineralized bone matrix. The present study describes an improved protocol for the isolation of human primary osteocytes. In contrast to an already established protocol, resting steps between the demineralization /digestion steps of the bone particles considerably improved the yield of osteocytes. Real-time polymerase chain reaction (PCR) analysis revealed the expression of typical osteocyte markers like osteocalcin, E11/podoplanin and dentin matrix protein 1 (DMP-1).

中文翻译：

---

一种从骨中分离原代人骨细胞的改进方法

骨细胞在骨代谢中非常重要，因为它们协调骨重塑、对机械感觉刺激作出反应并具有内分泌功能。体外因此，骨细胞研究与生物材料和药物测试具有高度相关性。使用原代人类细胞代替啮齿动物骨细胞系（如 MLOY4 和 IDG SW3）是可取的，但对分离这些深深嵌入矿化骨基质中的细胞提出了挑战。本研究描述了一种用于分离人类原代骨细胞的改进方案。与已经建立的方案相比，骨颗粒的脱矿质/消化步骤之间的休息步骤大大提高了骨细胞的产量。实时聚合酶链反应 (PCR) 分析揭示了典型骨细胞标志物如骨钙素、E11/podoplanin 和牙本质基质蛋白 1 (DMP-1) 的表达。