Characterization of the calcification process modeled in rat embryonic calvarial culture
Characterization of the calcification process modeled in rat embryonic calvarial culture
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DOI：
10.1093/jmicro/dfr068
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发表时间：
2011-10-01
期刊：
影响因子：
--
通讯作者：
Sasano, Yasuyuki
中科院分区：
文献类型：
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作者：
Kimura, Yasuko;Kikunaga, Shigeshi;Sasano, Yasuyuki
以大鼠胚胎颅骨为实验材料，设计了一种模拟生理性钙化过程的器官培养系统。从18天和20天（E18和E20）的大鼠胚胎中解剖标准化颅骨外植体，并培养1天、3天和5天。通过原子吸收分光光度法定量培养外植体的钙含量。将等同的外植体固定，包埋在石蜡中，切片并用von Kossa染色剂结合苏木精-伊红染色或进行能量色散X射线光谱处理，以确定组织中钙、磷和碳的浓度。培养5天后，E18和E20培养的颅骨（E18 cc和E20 cc）中总钙含量显著增加。所有培养的颅盖骨均呈von Kossa阳性，但在5天的培养期内，仅在E18 cc中观察到骨基质中的成骨细胞和骨细胞。钙和碳的浓度在E18 cc中在5天内显著增加，而E20几乎没有增加。E18 cc发生生理性钙化，E20 cc无。这些结果表明，使用E18颅骨的器官培养系统可用于体外模拟生理性钙化过程。
An organ culture system to model the physiological calcification process was designed using rat embryonic calvaria as a device for analyzing its mechanism. Standardized calvarial explants were dissected from rat embryos aged 18 and 20 days (E18 and E20) and cultured for 1, 3 and 5 days. The calcium content of the cultured explants was quantified by atomic absorption spectrophotometry. Equivalent explants were fixed, embedded in paraffin, sectioned and stained with von Kossa stain combined with hematoxylin-eosin or processed for energy-dispersive X-ray spectroscopy to determine the concentrations of calcium, phosphorus and carbon in the tissue. The total calcium content increased significantly in E18 and E20 cultured calvaria (E18cc and E20cc) over 5 days of culture. All cultured calvaria were von Kossa-positive, whereas the staining was intensified, and sound osteoblasts and osteocytes were observed in the bone matrix only in E18cc during the 5-day culture period. Concentrations of calcium and carbon increased significantly in E18cc over 5 days, whereas E20 showed little increase. Physiological calcification proceeded in E18cc, but not in E20cc. These results indicate that the organ culture system using E18 calvaria is useful for modeling the physiological calcification process in vitro.