Inhibition of growth and differentiation of osteoprogenitors in mouse bone marrow stromal cell cultures by increased donor age and glucocorticoid treatment
Inhibition of growth and differentiation of osteoprogenitors in mouse bone marrow stromal cell cultures by increased donor age and glucocorticoid treatment
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DOI：
10.1016/j.bone.2004.03.019
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发表时间：
2004-07-01
期刊：
影响因子：
4.1
通讯作者：
Chen, TL
中科院分区：
文献类型：
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作者：
Chen, TL
采用C57BL/6雄性小鼠幼龄（4-5月龄）和老龄（22-25月龄）长骨骨髓间质细胞（BMSC）原代培养，研究供体年龄对成骨细胞生长分化及对地塞米松（DEX）敏感性的影响。我们评估了碱性磷酸酶阳性骨形成成骨菌落（CFU-ALP）的数量和面积的变化以及包括ALP阴性菌落的菌落总数（CFU-F）的变化。测定细胞增殖、凋亡、ALP比活性，观察其生长分化情况。我们发现，从老年小鼠身上收获的有核细胞数量明显高于年轻小鼠（约多20%）。然而，在相同密度下，老年细胞形成的菌落数量和总面积小于年轻细胞形成的菌落。年轻细胞和年老细胞对DEX的反应相似，表现出集落数量和面积的剂量依赖性减少，面积的抑制大于数量的抑制。DEX对CFU-ALP的影响大于CFU-F，表明其对成骨细胞的抑制作用大于其他细胞类型。培养初期抑制细胞附着是导致DEX菌落数量减少的主要原因，抑制细胞增殖是导致面积减少的主要原因。溴脱氧尿苷（BrdU）掺入量严重呈剂量依赖性降低，低于对照组的40%。虽然DEX处理的培养物中凋亡细胞的数量较高，但凋亡并不是主要因素，因为即使DEX处理，凋亡细胞的数量也不到5%。尽管对集落数量和大小有这些负面影响，dex通过刺激集落ALP活性来增强成骨细胞分化活性，在年轻人中高达25倍，在老年人中高达5倍。我们的数据表明，年龄的增加降低了BMSC培养中骨祖细胞的数量及其生长。DEX降低了骨髓间充质干细胞的附着和增殖。这些变化反映了年龄相关性和糖皮质激素诱导的骨质减少。因此，小鼠骨髓间充质干细胞培养物可作为研究II型骨质疏松机制的有效体外模型。(C) 2004爱思唯尔公司版权所有。
Primary cultures of bone marrow stromal cells (BMSC) from long bones of young (4-5 months) and old (22-25 months) C57BL/6 male mice were used to study how donor age affects growth and differentiation of ostcoblasts and their sensitivity to dexamethasone (DEX). We assessed changes in the number and area of alkaline phosphatase-positive bone-forming osteolastic colonies (CFU-ALP) and in the total number of colonies (CFU-F) that include ALP negative colonies. Cell proliferation and apoptosis, specific activity of ALP, were also measured for growth and differentiation. We found that the number of nucleated cells harvested from old mice was significantly higher (approximately 20% more) than that from young mice. However, the number of colonies formed by old cells was fewer and the total area less than those formed by young cells plated at the same density. Young and old cells responded similarly to DEX showing a dose-dependent decrease in colony number and area with more inhibition for area than number. DEX affected CFU-ALP more than CFU-F indicating a greater inhibition for osteoprogenitor cells than other cell types. Inhibition of cell attachment at early culture was the major cause for the DEX reduction of colony number and the major cause of area reduction was inhibition of cell proliferation. This was demonstrated by a severe dose-dependent lowering of bromodeoxyuridine (BrdU) incorporation to less than 40% of the control. Although the number of apoptotic cells in the DEX-treated cultures was higher, apoptosis was not a major factor since the number of apoptotic cells was less than 5% even with DEX treatment. Despite these negative effects on colony number and size, DEX-enhanced osteoblastic differentiation activity by stimulating ALP activity of the colonies up to 25-fold in the young and 5-fold in the old. Our data suggest that increased age lowered the number of osteoprogenitor cells and their growth in BMSC cultures. DEX decreased the attachment and proliferation of BMSC in culture. These changes reflect age-related and glucocorticoid-induced osteopenia. Mouse BMSC cultures therefore may serve as a useful in vitro model to study the mechanisms of type II osteoporosis. (C) 2004 Elsevier Inc. All rights reserved.