Migration of culture-expanded human mesenchymal stem cells through bone marrow endothelium is regulated by matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-3
Migration of culture-expanded human mesenchymal stem cells through bone marrow endothelium is regulated by matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase-3
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DOI：
10.3324/haematol.10475
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发表时间：
2007-04-01
期刊：
影响因子：
10.1
通讯作者：
Van Riet, Ivan
中科院分区：
文献类型：
--
作者：
De Becker, Ann;Van Hummelen, Paul;Van Riet, Ivan
背景与目的骨髓间充质干细胞(MSC)是一种成体干细胞，可在体外扩增数倍，在造血干细胞移植的清髓处理后，具有恢复骨髓微环境和支持造血恢复的治疗潜力。成功地归巢到靶组织，如骨髓，意味着骨髓间充质干细胞在全身给药后能够外渗。然而，到目前为止，对MSC的外渗能力和潜在的机制还知之甚少。我们在体外研究了MSC通过骨髓内皮细胞的迁移能力。设计与方法进行了体外侵袭和跨内皮迁移实验。逆转录-聚合酶链式反应(RT-PCR)和酶谱法检测基质金属蛋白酶(MMP)的表达。比较高融合和低融合条件下培养细胞的迁移情况，并利用基因芯片和实时荧光定量RT-PCR分析不同条件下的差异基因表达。用中和抗MMP2抗体、MMP2短干扰RNA或重组金属蛋白酶组织抑制物(TIMP-3)检测MSC迁移的功能。结果MSC可侵袭重组基底膜，骨髓内皮细胞刺激这一过程。我们还发现MSC的跨内皮迁移至少部分受基质金属蛋白酶-2的调节。高度融合培养增加了天然基质金属蛋白酶抑制因子TIMP-3的产生，减少了骨髓基质细胞的跨内皮迁移。此外，文化融合程度对MSC的迁移也有显著影响。培养融合的增加阻碍了迁移，并与TIMP-3的上调有关。MSC的治疗应用将受益于对培养条件的选择，这些培养条件允许这些细胞的最佳外渗。
Background and ObjectivesMesenchymal stem cells (MSC) are adult stem cells that can be expanded many fold in vitro and have the therapeutic potential to restore the bone marrow microenvironment and support hematopoietic recovery after myeloablative conditioning for hematopoietic stem cell transplantation. Successful homing to the target tissue, such as bone marrow, implies that MSC are able to extravasate after systemic administration. However, the extravasation capacity of MSC and the underlying mechanisms are poorly understood to date. We studied in vitro the capacity of MSC to migrate through bone marrow endothelium.Design and MethodsIn vitro invasion and transendothelial migration assays were performed. The expression of matrix metalloproteinase (MMP) was analyzed by reverse transcriptase polymerase chain reaction (RT-PCR) and zymography. Migration of cells cultured at high or low confluence was compared and differential gene expression in these conditions was analyzed with microarray and real-time RT-PCR. The functional involvement in MSC migration was assessed using neutralizing anti-MMP-2 antibody, MMP-2 short interfering RNA or recombinant tissue inhibitor of metalloproteinase (TIMP-3).ResultsWe demonstrated that MSC can invade reconstituted basement membrane and that bone marrow endothelial cells stimulate this process. We also showed that the transendothelial migration of MSC is at least partially regulated by MMP-2. High culture confluence was found to increase production of the natural MMP-inhibitor TIMP-3 and decrease transendothelial migration of MSC.Interpretation and ConclusionsWe show that MSC have the potential to migrate through bone marrow endothelium and that this process involves MMP-2. Moreover, the migration of MSC is significantly influenced by the level of culture confluence. Increased culture confluence impairs migration and is related to an upregulation of TIMP-3. The therapeutic use of MSC would benefit from a selection of culture conditions that allow optimal extravasation of these cells.