Pereira Ana Rita S, Mendes Teresa F, Ministro Augusto, Teixeira Mariana, Filipe Mariana, Santos Jorge M, Bárcia Rita N, Goyri-O'Neill J, Pinto Fausto, Cruz Pedro E, Cruz Helder J, Santos Susana Constantino Rosa
Centro Cardiovascular da Universidade de Lisboa, Av. Prof Egas Moniz, 1649-028, Lisbon, Portugal.
Centro Hospitalar Lisboa Norte, Av. Prof. Egas Moniz, 1649-035, Lisbon, Portugal.
Stem Cell Res Ther. 2016 Sep 29;7(1):145. doi: 10.1186/s13287-016-0410-4.
Mesenchymal stem cells derived from human umbilical cord tissue, termed UCX®, have the potential to promote a full range of events leading to tissue regeneration and homeostasis. The main goal of this work was to investigate UCX® action in experimentally induced hindlimb ischemia (HLI).
UCX®, obtained by using a proprietary technology developed by ECBio (Amadora, Portugal), were delivered via intramuscular injection to C57BL/6 females after unilateral HLI induction. Perfusion recovery, capillary and collateral density increase were evaluated by laser doppler, CD31 immunohistochemistry and diaphonisation, respectively. The activation state of endothelial cells (ECs) was analysed after EC isolation by laser capture microdissection microscopy followed by RNA extraction, cDNA synthesis and quantitative RT-PCR analysis. The UCX®-conditioned medium was analysed on Gallios flow cytometer. The capacity of UCX® in promoting tubulogenesis and EC migration was assessed by matrigel tubule formation and wound-healing assay, respectively.
We demonstrated that UCX® enhance angiogenesis in vitro via a paracrine effect. Importantly, after HLI induction, UCX® improve blood perfusion by stimulating angiogenesis and arteriogenesis. This is achieved through a new mechanism in which durable and simultaneous upregulation of transforming growth factor β2, angiopoietin 2, fibroblast growth factor 2, and hepatocyte growth factor, in endothelial cells is induced by UCX®.
In conclusion, our data demonstrate that UCX® improve the angiogenic potency of endothelial cells in the murine ischemic limb suggesting the potential of UCX® as a new therapeutic tool for critical limb ischemia.
源自人脐带组织的间充质干细胞，称为UCX®，有促进一系列导致组织再生和内环境稳定事件的潜力。这项工作的主要目标是研究UCX®在实验性诱导的后肢缺血（HLI）中的作用。
通过使用ECBio（葡萄牙阿马多拉）开发的专有技术获得的UCX®，在单侧HLI诱导后经肌肉注射给予C57BL/6雌性小鼠。分别通过激光多普勒、CD31免疫组织化学和透明化评估灌注恢复、毛细血管和侧支密度增加情况。通过激光捕获显微切割显微镜分离内皮细胞（ECs），然后进行RNA提取、cDNA合成和定量RT-PCR分析，以分析ECs的激活状态。在Gallios流式细胞仪上分析UCX®条件培养基。分别通过基质胶小管形成和伤口愈合试验评估UCX®促进小管生成和EC迁移的能力。
我们证明UCX®通过旁分泌效应增强体外血管生成。重要的是，在HLI诱导后，UCX®通过刺激血管生成和动脉生成改善血液灌注。这是通过一种新机制实现的，其中UCX®诱导内皮细胞中转化生长因子β2、血管生成素2、成纤维细胞生长因子2和肝细胞生长因子的持久且同时上调。
总之，我们的数据表明UCX®提高了小鼠缺血肢体中内皮细胞的血管生成能力，提示UCX®作为严重肢体缺血新治疗工具的潜力。
