Replicative aging and gene expression in long-term cultures of human bone marrow stromal cells
Replicative aging and gene expression in long-term cultures of human bone marrow stromal cells
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DOI：
10.1089/107632702320934001
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发表时间：
2002-12-01
影响因子：
--
通讯作者：
Quarto, R
中科院分区：
文献类型：
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作者：
Banfi, A;Bianchi, G;Quarto, R
骨髓基质细胞（BMSCs）可以很容易地从成人骨髓中分离出来，并含有一群多能祖细胞，可以在体外和体内产生不同的间充质谱系。这些特性使得BMSCs成为一种有吸引力的靶点，用于各种疾病的基于细胞的治疗策略。然而，由于它们在体内的低频率，为了获得足够数量的细胞用于组织工程，需要广泛的体外扩增步骤，这可能显著改变BMSC的性质。因此，BMSCs的有效治疗用途需要设计合适的体外细胞扩增方法。在这项研究中，我们研究了体外扩增对BMSC增殖能力和自发分化的生物学效应。端粒酶活性和端粒缩短动力学与成骨，软骨形成和脂肪形成基因表达的变化在整个BMSC的寿命一起进行了评价。在培养中，骨髓间充质干细胞从未显示端粒酶活性，并且在体外扩增过程中端粒长度减少。此外，基因表达模式自发变化，在扩张过程中，表明人口向成骨谱系的渐进承诺。总之，培养的骨髓间充质干细胞进行复制老化和成骨分化。这些观察结果与其在临床上的成功使用有关，在设计新的治疗策略时应予以考虑。
Bone marrow stromal cells (BMSCs) can be easily isolated from adult marrow and contain a population of pluripotent progenitors that can give rise to different mesenchymal lineages both in vitro and in vivo. These properties make BMSCs an attractive target for cell-based therapeutic strategies for a variety of disorders. However, because of their low frequency in vivo, to obtain a sufficient number of cells for tissue engineering a step of extensive in vitro expansion is required, which could significantly alter BMSC properties. Therefore, effective therapeutic use of BMSCs requires the design of appropriate approaches for in vitro cell expansion. In this study we have investigated the biological effects of in vitro expansion on BMSC proliferative ability and on their spontaneous differentiation. Telomerase activity and telomere shortening kinetics were evaluated together with variations in osteogenic, chondrogenic, and adipogenic gene expression throughout the BMSC life span. In culture BMSCs never displayed telomerase activity and during in vitro expansion telomere length decreased. Furthermore, gene expression patterns spontaneously varied during expansion, indicating a progressive commitment of the population toward the osteogenic lineage. In conclusion, BMSCs in culture undergo progressive replicative aging and osteogenic differentiation. These observations are relevant to their successful use in clinics and should be considered when designing novel therapeutic strategies.