Murine marrow-derived mesenchymal stem cell:: isolation, in vitro expansion, and characterization
Murine marrow-derived mesenchymal stem cell:: isolation, in vitro expansion, and characterization
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DOI：
10.1046/j.1365-2141.2003.04669.x
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发表时间：
2003-11-01
影响因子：
6.5
通讯作者：
Nardi, NB
中科院分区：
文献类型：
--
作者：
Meirelles, LD;Nardi, NB
尽管对人类和其他物种骨髓 (BM) 来源的间充质干细胞 (MSC) 的研究给予了关注，但仍缺乏有关小鼠 MSC 的信息。我们描述了小鼠 BM 塑料贴壁细胞体外扩增超过 50 代的条件的建立，并提供了它们在形态、表面标记物轮廓和生长动力学方面的表征。这些细胞被证明沿着成骨和脂肪形成途径分化，并支持造血干细胞的生长和分化，因此在操作上被定义为鼠间充质干细胞（mMSC）。 mMSC 表面标志物 CD44、CD49e、CD29 和 Sca-1 呈阳性，并表现出均质、独特的形态。它们在成年 BALB/c 和 C57Bl/6 小鼠（α-L-艾杜糖醛酸酶（IDUA）基因正常或敲除）的 BM 中的频率初步估计为每 11 300-27 000 个有核细胞 1 个。 mMSC 培养的明确方法的出现，以及对其生物学的全面了解，将使小鼠模型中细胞和基因治疗方案的开发成为可能，并为成体干细胞研究领域提供新的视角。
In spite of the attention given to the study of mesenchymal stem cells (MSCs) derived from the bone marrow (BM) of humans and other species, there is a lack of information about murine MSCs. We describe the establishment of conditions for the in vitro expansion of plastic-adherent cells from murine BM for over 50 passages, and provide their characterization regarding morphology, surface marker profile and growth kinetics. These cells were shown to differentiate along osteogenic and adipogenic pathways, and to support the growth and differentiation of haematopoietic stem cells, and were thus operationally defined as murine mesenchymal stem cells (mMSCs). mMSCs were positive for the surface markers CD44, CD49e, CD29 and Sca-1, and exhibited a homogeneous, distinctive morphology. Their frequency in the BM of adult BALB/c and C57Bl/6 mice, normal or knockout for the alpha-L-iduronidase (IDUA) gene, was preliminarily estimated to be 1 per 11 300-27 000 nucleated cells. The emergence of a defined methodology for the culture of mMSCs, as well as a comprehensive understanding of their biology, will make the development of cellular and genetic therapy protocols in murine models possible, and provide new perspectives in the field of adult stem cells research.