Extracellular Vesicles from Vascular Endothelial Cells Promote Survival, Proliferation and Motility of Oligodendrocyte Precursor Cells.
Extracellular Vesicles from Vascular Endothelial Cells Promote Survival, Proliferation and Motility of Oligodendrocyte Precursor Cells.
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DOI：
10.1371/journal.pone.0159158
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发表时间：
2016
期刊：
影响因子：
3.7
通讯作者：
Ishizaki Y
中科院分区：
文献类型：
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作者：
Kurachi M;Mikuni M;Ishizaki Y
我们之前研究了脑微血管内皮细胞（MVEC）移植对大鼠白质梗死的影响，发现MVEC移植促进梗死区域脱髓鞘轴突的髓鞘再生，减少少突胶质细胞前体细胞（OPC）的凋亡。我们还发现培养的 MVEC 的条件培养基 (CM) 抑制培养的 OPC 的凋亡。在这项研究中，我们研究了 CM 中所含的细胞外囊泡 (EV) 对其 OPC 凋亡抑制作用的贡献。通过超速离心从 CM 中去除 EV 降低了其对 OPC 凋亡的抑制作用。为了确认源自 MVEC 的 EV 是否被培养的 OPC 吸收，我们用荧光染料 PKH67 标记 EV，并将其添加到 OPC 培养物中。添加后立即在 OPC 中发现许多用 PKH67 标记的囊泡结构。接下来我们研究了 MVEC 衍生的 EV 对 OPC 行为的影响。与 EV 一起培养 2 天后，与对照相比，固缩明显减少，BrdU 阳性 OPC 增多。我们还研究了 EV 对 OPC 运动的影响。与对照组相比，OPC 在 EV 存在的情况下迁移时间更长。为了检查内皮细胞的 EV 是否也对培养的 OPC 产生这些影响，我们从几种类型的内皮细胞的条件培养基中制备了 EV，并测试了它们对培养的 OPC 的影响。我们检查了来自所有类型内皮细胞的 EV 减少了 OPC 的凋亡并促进了它们的运动。鉴定内皮细胞 EV 中所含的分子可能有助于建立脱髓鞘疾病的有效疗法。
We previously examined the effect of brain microvascular endothelial cell (MVEC) transplantation on rat white matter infarction, and found that MVEC transplantation promoted remyelination of demyelinated axons in the infarct region and reduced apoptotic death of oligodendrocyte precursor cells (OPCs). We also found that the conditioned medium (CM) from cultured MVECs inhibited apoptosis of cultured OPCs. In this study, we examined contribution of extracellular vesicles (EVs) contained in the CM to its inhibitory effect on OPC apoptosis. Removal of EVs from the CM by ultracentrifugation reduced its inhibitory effect on OPC apoptosis. To confirm whether EVs derived from MVECs are taken up by cultured OPCs, we labeled EVs with PKH67, a fluorescent dye, and added them to OPC cultures. Many vesicular structures labeled with PKH67 were found within OPCs immediately after their addition. Next we examined the effect of MVEC-derived EVs on OPC behaviors. After 2 days in culture with EVs, there was significantly less pyknotic and more BrdU-positive OPCs when compared to control. We also examined the effect of EVs on motility of OPCs. OPCs migrated longer in the presence of EVs when compared to control. To examine whether these effects on cultured OPCs are shared by EVs from endothelial cells, we prepared EVs from conditioned media of several types of endothelial cells, and tested their effects on cultured OPCs. EVs from all types of endothelial cells we examined reduced apoptosis of OPCs and promoted their motility. Identification of the molecules contained in EVs from endothelial cells may prove helpful for establishment of effective therapies for demyelinating diseases.