Human bone cell cultures in biocompatibility testing.: Part I:: osteoblastic differentiation of serially passaged human bone marrow cells cultured in α-MEM and in DMEM
Human bone cell cultures in biocompatibility testing.: Part I:: osteoblastic differentiation of serially passaged human bone marrow cells cultured in α-MEM and in DMEM
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DOI：
10.1016/s0142-9612(99)00284-7
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发表时间：
2000-06-01
期刊：
影响因子：
14
通讯作者：
Fernandes, MH
中科院分区：
文献类型：
--
作者：
Coelho, MJ;Cabral, AT;Fernandes, MH
良好表征的人成骨细胞骨髓细胞培养物是分析骨组织/生物材料相互作用的有用体外工具。在这项工作中，在实验条件下培养人骨髓，以促进成骨细胞分化，并在两种广泛使用的培养基中培养连续传代的细胞，最低必需培养基Eagle，α改良（α-MEM）和Dulbecco改良Eagle培养基（DMEM）。培养物生长35 d，并比较扫描电子显微镜（SEM）上的形态学外观、细胞活力/增殖、总蛋白含量、碱性磷酸酶（ALP）活性和形成磷酸钙沉积物的能力。结果表明，在两种培养基中生长的培养物中细胞增殖相似，但在DMEM培养物中ALP活性和形成矿化沉积物的能力较低。在这两种实验情况下，成骨细胞参数强烈降低细胞传代，特别是从第一次到第二次传代培养。在使用的实验条件下（原代和传代培养物中存在抗坏血酸、β-甘油磷酸钠和地塞米松），在α-MEM中生长的第一次和第二次传代培养物以及在DMEM中生长的第一次传代培养物中观察到成骨细胞分化。这些结果强调了在涉及骨细胞培养的研究中定义实验条件的重要性。(C)2000爱思唯尔科技有限公司版权所有。
Well-characterised human osteoblastic bone marrow cell cultures are a useful in vitro tool to analyse bone tissue/biomaterials interactions. In this work, human bone marrow was cultured in experimental conditions described to favour osteoblastic differentiation and, serially passaged cells were cultured in two widely used culture media, minimum essential medium Eagle, alpha modification (alpha-MEM) and Dulbecco's modified Eagle's medium (DMEM). Cultures were grown for 35 d and compared concerning morphologic appearance on scanning electron microscopy (SEM), cell viability/proliferation, total protein content, activity of alkaline phosphatase (ALP) and ability to form calcium phosphate deposits. Results showed that cell proliferation was similar in cultures grown in the two media but ALP activity and ability to form mineralised deposits were lower in DMEM cultures. In both experimental situations, osteoblastic parameters were strongly reduced on cell passage, particularly from the first to the second subculture. In the experimental conditions used (presence of ascorbic acid, sodium beta-glycerophosphate and dexamethasone in the primary and secondary cultures), osteoblastic differentiation was observed in the first and second subcultures grown in alpha-MEM and in the first subculture grown in DMEM. These results underline the importance of the definition of the experimental conditions in studies involving bone cell cultures. (C) 2000 Elsevier Science Ltd. All rights reserved.