Maintenance of osteoblastic and adipocytic differentiation potential with age and osteoporosis in human marrow stromal cell cultures
Maintenance of osteoblastic and adipocytic differentiation potential with age and osteoporosis in human marrow stromal cell cultures
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DOI：
10.1007/s00223-001-2059-x
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发表时间：
2002-07-01
影响因子：
4.2
通讯作者：
Kassem, M
中科院分区：
文献类型：
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作者：
Justesen, J;Stenderup, K;Kassem, M
成骨细胞和脂肪细胞在骨髓基质中有一个共同的前体细胞，称为骨髓基质细胞（MSC）。由于体内骨脂肪组织的体积随着年龄的增长而增加，我们假设在衰老期间和骨质疏松症（OP）患者中观察到的骨形成减少是MSC的脂肪生成增强和成骨细胞生成减少的结果。因此，从年轻供体（年龄18-42，n = 34）、老年健康供体（年龄66-78，n = 20）和OP患者（年龄58 - 76，n = 15）建立MSC培养物。将细胞在成脂培养基（含有15%马血清和100 nM地塞米松）、成骨培养基（含有10%胎牛血清[FCS]和10 nM骨化三醇）或对照培养基（10% FCS）中培养2周。通过其形成集落的能力鉴定MSC。对菌落总数、碱性磷酸酶染色阳性的成骨细胞菌落（AP（+））和含脂肪细胞的脂肪细胞菌落（Ad（+））进行定量。此外，使用逆转录聚合酶链反应（RTPCR）测定脂肪细胞和成骨细胞表型的基因标志物的稳态mRNA水平。成脂和成骨培养基分别诱导细胞分化和脂肪细胞和成骨细胞谱系特异性标志物的表达。我们发现成骨细胞或脂肪细胞集落形成或成脂或成骨基因标记物的mRNA的稳态水平没有年龄相关的变化。从OP患者中获得的细胞显示出与年龄匹配的对照组相似的分化模式。总之，MSC在衰老过程中和OP患者中保持其分化潜力。需要确定与年龄相关的骨形成减少的其他机制。
Osteoblasts and adipocytes share a common precursor cell in the bone marrow stroma, termed marrow stromal cell (MSC). As the volume of bone adipose tissue increases in vivo with age, we hypothesized that decreased bone formation observed during aging and in patients with osteoporosis (OP) is the result of enhanced adipogenesis and decreased osteoblastogenesis from the MSCs. Thus, cultures of MSCs were established from young donors (age 18-42, n = 34), elderly healthy donors (age 66-78, n 20), and patients with OP (age 5876, n = 15). Cells were cultured for 2 weeks in an adipogenic medium (containing 15% horse serum and 100 nM dexamethasone), osteogenic medium (containing 10% fetal calf serum [FCS] and 10 nM calcitriol), or control medium (10% FCS). The MSCs were identified by their abilities to form colonies. Total number of colonies, osteoblastic colonies stained positive for alkaline phosphatase (AP(+)), and adipocytic colonies containing adipocytes (Ad(+)) were quantitated. In addition, steady state mRNA levels of gene markers of adipocytic and osteoblastic phenotypes were determined using reverse-transcriptase polymerase chain reaction (RTPCR). The adipogenic and osteogenic media induced cell differentiation and the expression of adipocytic and osteoblastic lineage-specific markers, respectively. We found no age-related changes in the osteoblastic or adipocytic colony formation or the steady state levels of mRNA of the adipogenic or osteogenic gene markers. Cells obtained from patients with OP showed a pattern of differentiation similar to those of age-matched controls. In conclusion, MSCs maintain their differentiation potential during aging and in patients with OP. Other mechanisms responsible for age-related decrease in bone formation need to be determined.