INSERM, U1026, F-33000, Bordeaux, France.
J Tissue Eng Regen Med. 2012 Nov;6(10):e51-60. doi: 10.1002/term.1539. Epub 2012 Jun 28.
Association of the bone-forming osteoblasts (OBs) and vascular endothelial cells (ECs) into a biomaterial composite provides a live bone graft substitute that can repair the bone defect when implanted. An intimate functional relationship exists between these cell types. This communication is crucial to the coordinated cell behaviour necessary for bone development and remodelling. Previous studies have shown that direct co-culture of primary human osteoprogenitors (HOPs) with primary human umbilical vein endothelial cells (HUVECs) stimulates HOPs differentiation and induces tubular-like networks. The present work aims to test the use of human bone marrow stromal cells (HBMSCs) co-cultured with human endothelial progenitor cells in order to assess whether progenitor-derived ECs (PDECs) could support osteoblastic differentiation as mature ECs do. Indeed, data generated from the literature by different laboratories considering these co-culture systems appear difficult to compare. Monocultures of HUVECs, HOPs, HBMSCs (in a non-orientated lineage), PDECs (from cord blood) were used as controls and four combinations of co-cultures were undertaken: HBMSCs-PDECs, HBMSCs-HUVECs, HOPs-PDECs, HOPs-HUVECs with ECs (mature or progenitor) for 6 h to 7 days. At the end of the chosen co-culture time, intracellular alkaline phosphatase (ALP) activity was detected in HOPs and HBMSCs and quantified in cell extracts. Quantitative real-time polymerase chain reaction (qPCR) of ALP was performed over time and vascular endothelial growth factor (VEGF) was measured. After 21 days, calcium deposition was observed, comparing mono- and co-cultures. We confirm that ECs induce osteoblastic differentiation of mesenchymal stem cells in vitro. Moreover, HUVECs can be replaced by PDECs, the latter being of great interest in tissue engineering.
成骨细胞（OBs）和血管内皮细胞（ECs）与生物材料复合形成活骨移植物替代物，植入后可修复骨缺损。这些细胞类型之间存在密切的功能关系。这种通讯对于骨发育和重塑所需的协调细胞行为至关重要。先前的研究表明，将原代人成骨细胞（HOPs）与原代人脐静脉内皮细胞（HUVECs）直接共培养可刺激 HOPs 分化并诱导管状网络。本研究旨在测试共培养人骨髓基质细胞（HBMSCs）和人内皮祖细胞（PDECs），以评估祖细胞衍生的 ECs（PDECs）是否可以支持成骨细胞分化，就像成熟的 ECs 一样。事实上，不同实验室从文献中生成的数据考虑到这些共培养系统，似乎难以比较。单独培养 HUVECs、HOPs、HBMSCs（非定向谱系）、PDECs（来自脐带血）作为对照，并进行了四种共培养组合：HBMSCs-PDECs、HBMSCs-HUVECs、HOPs-PDECs、HOPs-HUVECs，与 ECs（成熟或祖细胞）共培养 6 小时至 7 天。在选定的共培养时间结束时，检测 HOPs 和 HBMSCs 中的细胞内碱性磷酸酶（ALP）活性，并在细胞提取物中定量。随着时间的推移，对 ALP 进行实时定量聚合酶链反应（qPCR），并测量血管内皮生长因子（VEGF）。21 天后，比较了单核和共培养物的钙沉积。我们证实 ECs 在体外诱导间充质干细胞的成骨细胞分化。此外，PDECs 可以替代 HUVECs，后者在组织工程中具有重要意义。
