Selection of common markers for bone marrow stromal cells from various bones using real-time RT-PCR: effects of passage number and donor age.

Akira Igarashi, Kazumi Segoshi, Yuhiro Sakai, Haiou Pan, Masami Kanawa, Yukihito Higashi, Masaru Sugiyama, Kozo Nakamura, Hidemi Kurihara, Satoru Yamaguchi, Koichiro Tsuji, Takeshi Kawamoto, Yukio Kato
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引用次数: 48

Abstract

Bone marrow stromal cells (BMSCs) are valuable in tissue engineering and cell therapy, but the quality of the cells is critical for the efficacy of therapy. To test the quality and identity of transplantable cells, we identified the molecular markers that were expressed at higher levels in BMSCs than in fibroblasts. Using numerous BMSC lines from tibia, femur, ilium, and jaw, together with skin and gum fibroblasts, we compared the gene expression profiles of these cells using DNA microarrays and low-density array cards. The differentiation potential of tibia and femur BMSCs was similar to that of iliac BMSCs, and different from jaw BMSCs, but all BMSC lines had many common markers that were expressed at much higher levels in BMSCs than in fibroblasts; several BMSC markers showed discrete expression patterns between jaw and other BMSCs. The common markers are probably useful in routine tests, but their efficacy may depend upon the passage number or donor age. In our study the passage number markedly altered the expression levels of several markers, while donor age had little effect on them. Considering the effects of in vivo location of BMSCs and passage, magnitude of increase in expression levels, and interindividual differences, we identified several reliable markers -- LIF, IGF1, PRG1, MGP, BMP4, CTGF, KCTD12, IGFBP7, TRIB2, and DYNC1I1 -- among many candidates. This marker set may be useful in a routine test for BMSCs in tissue engineering and cell therapy.

利用实时RT-PCR选择不同骨源骨髓基质细胞的共同标记物:传代数和供体年龄的影响
骨髓基质细胞(BMSCs)在组织工程和细胞治疗中具有重要的应用价值,但其质量是影响治疗效果的关键因素。为了测试可移植细胞的质量和特性,我们鉴定了在骨髓间充质干细胞中表达水平高于成纤维细胞的分子标记。利用来自胫骨、股骨、髂骨和颌骨的大量骨髓间充质干细胞系,以及皮肤和牙龈成纤维细胞,我们使用DNA微阵列和低密度阵列卡比较了这些细胞的基因表达谱。胫骨和股骨骨髓间充质干细胞的分化潜力与髂骨髓间充质干细胞相似,与颌骨骨髓间充质干细胞不同,但所有骨髓间充质干细胞系都有许多共同的标记物,这些标记物在骨髓间充质干细胞中的表达水平远高于成纤维细胞;一些骨髓间充质干细胞标记物在颌骨和其他骨髓间充质干细胞之间表现出离散的表达模式。普通标记可能在常规检查中有用,但其功效可能取决于传代次数或供体年龄。在我们的研究中,传代数显著改变了几种标志物的表达水平,而供体年龄对它们的影响很小。考虑到骨髓间充质干细胞在体内的位置和传代、表达水平增加的幅度以及个体间差异的影响,我们在许多候选物中确定了几个可靠的标记物——LIF、IGF1、PRG1、MGP、BMP4、CTGF、KCTD12、IGFBP7、TRIB2和DYNC1I1。该标记集可用于组织工程和细胞治疗中骨髓间充质干细胞的常规检测。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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来源期刊
Tissue engineering
Tissue engineering CELL & TISSUE ENGINEERING-BIOTECHNOLOGY & APPLIED MICROBIOLOGY
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