Tropel Philippe, Noël Danièle, Platet Nadine, Legrand Pierre, Benabid Alim-Louis, Berger François
INSERM U318, Grenoble, France.
Exp Cell Res. 2004 May 1;295(2):395-406. doi: 10.1016/j.yexcr.2003.12.030.
The future use of adult mesenchymal stem cells (MSCs) for human therapies depends on the establishment of preclinical studies with other mammals such as mouse. Surprisingly, purification and characterisation of murine MSCs were only poorly documented. The aim of this study was to purify mouse MSCs from adult bone marrow and to functionally characterise their abilities to differentiate along diverse lineages. Adherent cells from adult C57Bl/6J mouse bone marrow were depleted of granulo-monocytic cells and subsequently allowed to grow on fibronectin-coated dishes in presence of fetal bovine serum and growth factors. The growing fibroblastoid cell population primarily consisted of spindle- and star-shaped cells with significant renewal capacity as they were cultured until 30 passages (about 60 doubling population). We fully demonstrated the MSC phenotype of these cells by inducing them to differentiate along osteoblastic, adipocytic, and chondrocytic pathways. Mouse MSCs (mMSCs) sharing the same morphological and functional characteristics as human MSCs can be successfully isolated from adult bone marrow without previous mouse or bone marrow treatment. Therefore, mMSCs will be an important tool to study the in vivo behaviour and fate of this cell type after grafting in mouse pathology models.
成人间充质干细胞(MSCs)在人类治疗中的未来应用取决于与其他哺乳动物(如小鼠)进行临床前研究。令人惊讶的是,关于小鼠MSCs的纯化和特性描述记录很少。本研究的目的是从成年骨髓中纯化小鼠MSCs,并对其沿不同谱系分化的能力进行功能特性分析。从成年C57Bl/6J小鼠骨髓中分离出的贴壁细胞去除粒单核细胞,随后在存在胎牛血清和生长因子的情况下,使其在纤连蛋白包被的培养皿上生长。生长的成纤维样细胞群体主要由纺锤形和星形细胞组成,具有显著的更新能力,因为它们被培养至30代(约60次群体倍增)。通过诱导这些细胞沿成骨、脂肪生成和软骨生成途径分化,我们充分证明了这些细胞的MSC表型。与人类MSCs具有相同形态和功能特征的小鼠MSCs(mMSCs)可以成功地从成年骨髓中分离出来,无需事先对小鼠或骨髓进行处理。因此,mMSCs将成为研究这种细胞类型在小鼠病理模型中移植后体内行为和命运的重要工具。