Mesenchymal stem cells transmigrate across brain microvascular endothelial cell monolayers through transiently formed inter-endothelial gaps
Mesenchymal stem cells transmigrate across brain microvascular endothelial cell monolayers through transiently formed inter-endothelial gaps
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DOI：
10.1016/j.neulet.2011.07.021
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发表时间：
2011-09-08
影响因子：
2.5
通讯作者：
Shimohama, Shun
中科院分区：
文献类型：
--
作者：
Matsushita, Takashi;Kibayashi, Tatsuya;Shimohama, Shun
间充质干细胞(MSCs)在脑缺血、帕金森病等神经系统疾病的细胞治疗中具有广阔的应用前景。静脉注射间充质干细胞在脑实质内积聚，但对骨髓间充质干细胞跨血脑屏障(BBB)迁移的细节知之甚少。为了研究MSC跨血脑屏障的迁移，我们建立了一个由大鼠脑微血管内皮细胞(BMECs)和骨髓来源的MSCs组成的体外培养系统，使用Transwell或Millicell培养块。利用该系统，我们首先研究了上腔中加入的MSCs数量对BMEC屏障完整性的影响。加入密度为1.5×10(5)cell/cm(2)的MSCs可破坏BMEC的单层结构，并通过跨内皮细胞电阻(TEER)测定屏障功能。当细胞密度为1.5×10~(4)个/cm~(2)时，在此条件下培养24小时后，BMEC单层未见明显破坏，TEER也未见明显下降。在内皮下间隙或植入膜下可见MSCs，提示MSCs可跨BMEC单层迁移。时间推移成像显示，MSCs通过在BMEC之间瞬时形成的细胞间隙跨BMEC单层迁移。这些结果表明，我们建立的由骨髓间充质干细胞和骨髓间充质干细胞组成的体外培养系统对于研究骨髓间充质干细胞跨血脑屏障迁移的分子和细胞机制是有用的。(C)2011爱思唯尔爱尔兰有限公司。保留所有权利。
Mesenchymal stem cells (MSCs) hold much promise for cell therapy for neurological diseases such as cerebral ischemia and Parkinson's disease. Intravenously administered MSCs accumulate in lesions within the brain parenchyma, but little is known of the details of MSC transmigration across the blood-brain barrier (BBB). To study MSC transmigration across the BBB, we developed an in vitro culture system consisting of rat brain microvascular endothelial cells (BMECs) and bone marrow-derived MSCs using Transwell or Millicell culture inserts. Using this system, we first investigated the influence of the number of MSCs added to the upper chamber on BMEC barrier integrity. The addition of MSCs at a density of 1.5 x 10(5) cells/cm(2) led to disruption of the BMEC monolayer structure and decreased barrier function as measured by the transendothelial electrical resistance (TEER). When applied at a density of 1.5 x 10(4) cells/cm(2), neither remarkable disruption of the BMEC monolayers nor a significant decrease in TEER was observed until at least 12 h. After cultivation for 24 h under this condition. MSCs were found in the subendothelial space or beneath the insert membrane, suggesting that MSCs transmigrate across BMEC monolayers. Time-lapse imaging revealed that MSCs transmigrated across the BMEC monolayers through transiently formed intercellular gaps between the BMECs. These results show that our in vitro culture system consisting of BMECs and MSCs is useful for investigating the molecular and cellular mechanisms underlying MSC transmigration across the BBB. (C) 2011 Elsevier Ireland Ltd. All rights reserved.