Vu Duc M, Masuda Haruchika, Yokoyama Tun A, Fujimura Satoshi, Kobori Michiru, Ito Rie, Sawada Kaichiro, Saito Akira, Asahara Takayuki
Division of Nephrology and Metabolism, Department of Medicine, Tokai University School of Medicine, Kanagawa, Japan.
Tissue Eng Part A. 2009 Oct;15(10):3173-82. doi: 10.1089/ten.TEA.2009.0050.
Development of a bioartificial glomerulus, a hemofilter in which the inner surface of hollow fibers is endothelialized, requires expandable, nonimmunogenic, antithrombogenic, and highly permeable endothelial cells. We used human umbilical cord blood CD133(+) endothelial progenitor cells (EPCs) to evaluate the feasibility of application of EPCs for bioartificial glomerulus. Numbers of adhered CD133(+) EPCs (adhered EPCs) was approximately 25 to 30 times as great in the expansion culture group as in the non-expansion group. Adhered EPCs had endothelial cell features, including the expression of CD31, Kinase domain region, von Willebrand factor, vascular endothelial-cadherin, positive for Ulex europeus agglutinin I staining, and up-take of acetylated low-density lipoprotein. Adhered EPCs secreted 6-keto-prostaglandin F(1alpha) identically to that secreted by human umbilical vein endothelial cells (HUVECs). The cells also expressed messenger RNA for phospholipase A(2), cyclooxygenase (COX)-1, COX-2, prostaglandin I(2) synthase, tissue plasminogen activator, and thrombomodulin (TM). TM protein in adhered EPCs properly activated protein C. Scanning electron microscopy revealed the suppression of platelet adhesion and aggregation on the surface of cell monolayer. Adhered EPCs treated with 50 microg/mL of cytochalasin B induced a larger diameter and a greater number of fenestrae, subsequently producing significantly more ultrafiltration than the non-treated cell. These results suggest that CD133(+) EPCs would potentially be applicable in bioartificial glomerulus.
生物人工肾小球是一种中空纤维内表面内皮化的血液滤过器，其构建需要可扩增、无免疫原性、抗血栓形成且具有高通透性的内皮细胞。我们使用人脐带血CD133(+)内皮祖细胞（EPCs）来评估EPCs应用于生物人工肾小球的可行性。在扩增培养组中，贴壁的CD133(+) EPCs（贴壁EPCs）数量大约是非扩增组的25至30倍。贴壁EPCs具有内皮细胞特征，包括CD31、激酶结构域区域、血管性血友病因子、血管内皮钙黏蛋白的表达，荆豆凝集素I染色呈阳性，以及摄取乙酰化低密度脂蛋白。贴壁EPCs分泌6-酮-前列腺素F(1α)的情况与脐静脉内皮细胞（HUVECs）相同。这些细胞还表达磷脂酶A(2)、环氧化酶（COX）-1、COX-2、前列腺素I(2)合成酶、组织型纤溶酶原激活剂和血栓调节蛋白（TM）的信使RNA。贴壁EPCs中的TM蛋白能正常激活蛋白C。扫描电子显微镜显示细胞单层表面血小板黏附和聚集受到抑制。用50μg/mL细胞松弛素B处理的贴壁EPCs诱导出更大的直径和更多的窗孔，随后产生的超滤量明显多于未处理的细胞。这些结果表明CD133(+) EPCs可能适用于生物人工肾小球。
