In vitro bone formation by rat marrow cell culture
In vitro bone formation by rat marrow cell culture
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DOI：
10.1002/(sici)1097-4636(199611)32:3
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发表时间：
1996-11-01
期刊：
影响因子：
--
通讯作者：
Suwa, Y
中科院分区：
文献类型：
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作者：
Ohgushi, H;Dohi, Y;Suwa, Y
从Fischer大鼠股骨中获取新鲜骨髓细胞，在含有15%胎牛血清（FCS）的培养基中培养至浸出汇合。胰蛋白酶消化后，在FCS、10mM P-甘油磷酸盐、82μg/mL抗坏血酸磷酸盐和10(-8)M地塞米松(Dex)存在下，以100×10(3)/35mm孔的细胞密度传代培养细胞。继代培养后约1周开始出现成骨细胞和微观矿化结节，2周时许多肉眼可见的结节表现出高碱性磷酸酶活性（ALP）和骨Gla蛋白（BGP）mRNA的出现是明显的。原位杂交证明，长方体细胞（成骨细胞）有mRNA表现。 X 射线衍射 (XRD) 和傅里叶变换红外光谱 (FTIR) 显示羟基磷灰石细晶的矿化程度与天然大鼠骨矿物质相当。与这些发现相反，在相同条件下进行的传代培养，除了缺乏 Dex 外，没有显示矿化结节，也没有显示成骨细胞表型表达。这些分析表明，使用大鼠骨髓细胞培养物诱导的 Dex 矿化是体内骨形成的体外对应物。这种培养物可用于研究材料/成骨细胞的相互作用。 (C) 1996 约翰威利父子公司
Fresh marrow cells were obtained from the femora Fischer rats and cultured in a medium containing 15% fetal calf serum (FCS) to leach confluent. After trypsinization, cells were subcultured at a cell density of 100 x 10(3)/35 mm well in the presence of FCS, 10 mM P-glycerophosphate, 82 mu g/mL ascorbic acid phosphate, and 10(-8)M dexamethasone (Dex). Osteoblastic cells and microscopic mineralized nodules began to appear at about 1 week after the subculture, and at 2 weeks many macroscopic nodules that showed high alkaline phosphatase activity (ALP) and appearance of bone Gla protein (BGP) mRNA were evident. As demonstrated by in situ hybridization, the mRNA was manifested by cuboid-shaped cells (osteoblastic cells). X-ray diffraction (XRD) and Fourier-transform infrared spectroscopy (FTIR) showed the mineralization of fine crystals of hydroxyapatite comparable to natural rat bone mineral. In contrast to these findings, subculture done under the same conditions except for the lack of Dex did not show mineralized nodules, nor did they show the osteoblastic phenotype expression. These analyses indicate that Dex-induced mineralization using rat bone marrow cell culture is an in vitro counterpart of bone formed in vivo. Such a culture is useful for investigating materials/osteogenic cells interactions. (C) 1996 John Wiley & Sons, Inc.