| Binding to Secreted Bone Matrix in vitro
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Author:
Date:
2014-02-20
[Abstract] This method examines the bone matrix binding capacity of proteins. Using osteogenic differentiation medium, multipotent stromal cells (MSC) are induced to differentiate into osteocytes in vitro and to secrete bone matrix. The latter is confirmed using Alizarin red S staining, which detects the presence of calcific deposits (hydroxyapatite). These calcific deposits are used to test the bone binding properties of proteins. The binding to the calcific deposits is assessed by Western blot analysis.
[摘要] 这种方法检查蛋白质的骨基质结合能力。 使用成骨分化培养基，诱导多潜能基质细胞（MSC）在体外分化成骨细胞和分泌骨基质。 后者使用茜素红S染色证实，其检测钙化沉积物（羟基磷灰石）的存在。 这些钙沉积物用于测试蛋白质的骨结合性质。 通过蛋白质印迹分析评估与钙化沉积物的结合。
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| In vitro Biomineralization Assay
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Author:
Date:
2014-02-05
[Abstract] Biomineralization in vertebrates has both physiological and pathological aspects. Physiological mineralization is essential for proper development and function of hard tissues, such as bone, teeth, and growth plate cartilage, but it does not occur in soft tissues. Pathological ectopic mineralization, in contrast, occurs in soft tissues, including blood vessels, kidney, articular cartilage, and cardiovascular tissue. Here, we describe the simple method for detecting and measuring the presence of mineralized nodules in cardiac ventricular fibroblasts by using von Kossa and alizarin red S staining, and a colorimetric method for calcium quantification, respectively.
[摘要] 脊椎动物中的生物矿化具有生理和病理学方面。 生理矿化对于硬组织（例如骨，牙齿和生长板软骨）的正常发育和功能是必需的，但它不在软组织中发生。 相反，病理异位矿化发生在软组织中，包括血管，肾，关节软骨和心血管组织。 在这里，我们描述了使用von Kossa和茜素红S染色和钙定量的比色法分别检测和测量心室成纤维细胞中矿化结节的存在的简单方法。
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| Osteogenic and Adipogenic Differentiation of Osteosarcoma Cells
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Author:
Date:
2012-12-20
[Abstract] Osteosarcomas are the most common primary, non-hematologic malignant tumors in childhood and adolescence, comprising almost 60% of all bone sarcomas. Although these tumors are osteoblastic in nature, the characteristics of the specific tumor-initiating cells are unclear. Osteosarcomas contain highly proliferative undifferentiated malignant cells with a disrupted bone differentiation program. Cancer stem cells (CSCs) that have tumor-initiating properties and capacity of symmetric and asymmetric division have now been described in many solid tumors. For osteosarcomas, the CSC hypothesis has received support from recently reported findings that both human and murine osteosarcomas contain a sub-population of multipotent cells that that express various mesenchymal stem cell surface markers and ...
[摘要] 骨肉瘤是儿童和青少年中最常见的原发性非血液恶性肿瘤，占所有骨肉瘤的近60％。虽然这些肿瘤在性质上是成骨细胞的，但是特异性肿瘤起始细胞的特征不清楚。骨肉瘤包含具有破坏的骨分化程序的高度增殖性未分化的恶性细胞。已经在许多实体瘤中描述了具有肿瘤起始性质和对称和不对称分裂能力的癌干细胞（CSC）。对于骨肉瘤，CSC假说已经从最近报道的发现获得支持，即人和鼠骨肉瘤包含表达多种间充质干细胞表面标志物的多能细胞亚群，并且能够在多种间充质谱系例如成骨细胞和脂肪细胞。通过在特定培养基中生长细胞并测定分化标记物，可以容易地评估分化为这些不同谱系。
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