Therapeutic Angiogenesis by Implantation of a Capillary Structure Constituted of Human Adipose Tissue Microvascular Endothelial Cells
Therapeutic Angiogenesis by Implantation of a Capillary Structure Constituted of Human Adipose Tissue Microvascular Endothelial Cells
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通过植入由人脂肪组织微血管内皮细胞构成的毛细血管结构来治疗性血管生成
DOI：
10.1161/atvbaha.109.198994
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发表时间：
2010
期刊：
影响因子：
--
通讯作者：
M.Abe
中科院分区：
文献类型：
--
作者：
T.Yoshida;M.Komaki;H.Hattori;J.Negishi;A.Kishida;I.Morita;M.Abe
目的——我们之前报道了一种使用光学光刻技术进行毛细管网络工程的新技术。为了将该技术应用于缺血性疾病的治疗，我们测试了人网膜微血管内皮细胞（HOMEC）作为自体细胞来源和脱细胞人羊膜（DC-AM）作为无病原体和低免疫原性移植支架。方法和结果——人脐静脉内皮细胞在图案玻璃基板上排列，并在转移到羊膜（AM）上时形成毛细血管结构。相比之下，HOMECs 是分散的，并且没有在 AMs 上形成毛细管结构。用 1-磷酸鞘氨醇 (S1P) 处理 HOMEC 可抑制 HOMEC 迁移，并使 HOMEC 在 AM 上形成毛细管结构。通过定量 RT-PCR 和蛋白质印迹分析，我们证明 HOMEC 中的主要 S1P 受体是 S1P2，而人脐静脉内皮细胞中缺乏 S1P2，并且 S1P 对细胞迁移的抑制是通过 S1P2-Rho-Rho 相关激酶信号通路介导的。将 DC-AM 上工程化的毛细血管植入后肢缺血裸鼠模型中，与对照组相比，显着增加了血液灌注。 结论：可以使用打印技术和 S1P 处理离体工程设计由 DC-AM 上的 HOMEC 组成的毛细血管网络。这种毛细血管网络再生方法可能具有治疗缺血性疾病的潜力。
Objective—We previously reported a novel technology for the engineering of a capillary network using an optical lithographic technique. To apply this technology to the therapy of ischemic diseases, we tested human omental microvascular endothelial cells (HOMECs) as an autologous cell source and decellularized human amniotic membranes (DC-AMs) as a pathogen-free and low immunogenic transplantation scaffold.Methods and Results—Human umbilical vein endothelial cells were aligned on a patterned glass substrate and formed a capillary structure when transferred onto an amniotic membrane (AM). In contrast, HOMECs were scattered and did not form a capillary structure on AMs. Treatment of HOMECs with sphingosine 1-phosphate (S1P) inhibited HOMEC migration and enabled HOMEC formation of a capillary structure on AMs. Using quantitative RT-PCR and Western blot analyses, we demonstrated that the main S1P receptor in HOMECs is S1P2, which is lacking in human umbilical vein endothelial cells, and that inhibition of cell migration by S1P is mediated through an S1P2–Rho–Rho-associated kinase signaling pathway. Implantation of capillaries engineered on DC-AMs into a hindlimb ischemic nude mouse model significantly increased blood perfusion compared with controls.Conclusion—A capillary network consisting of HOMECs on DC-AMs can be engineered ex vivo using printing technology and S1P treatment. This method for regeneration of a capillary network may have therapeutic potential for ischemic diseases.