ISOLATION OF ENDOTHELIAL PROGENITOR CELL FROM HUMAN UMBILICAL CORD BLOOD

从人脐带血中分离内皮祖细胞

• 批准号: 11557058
• 负责人: MUROHARA Toyoaki
• 金额: $ 8.64万
• 依托单位: KURUME UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11557058/
• 关键词: ENDOTHELIAL PROGENITOR CELLS; ISCHEMIC HEART DISEASE; VASCULAR REGENERATION; PERIPHERAL ARTERY OCCLUSIVE DISEASE; VASCULOGENESIS; ANGIOGENESIS; ENDOTHELIAL CELL; COLLATERAL VESSEL; 再生医学; 幹細胞; 動脈硬化; 臍帯血; 成人末梢血

Endothelial precursor cells (EPCs) have been identified in adult peripheral blood. We examined whether EPCs could be isolated from Umbilical cord blood, a rich source for hematopoietic progenitors, and whether in vivo transplantation of EPCs could modulate postnatal neovascularization. Numerous cell clusters, spindle-shaped and attaching (AT) cells, and cord-like structures developed from culture of cord blood mononuclear cells (MNCs). Fluorescence-trace experiments revealed that cell clusters, AT cells and cord-like structures predominantly derived from CD34-positive MNCs (MNC^<CD34+>). Greater numbers of AT cells and cell clusters developed from cord blood-MNCs than from an equal amount of adult peripheral blood-MNCs. AT cells incorporated acetylated-LDL, released nitric oxide, and expressed KDR, VE-cadherin, CD31, and von Willebrand factor but not CD45. Locally transplanted AT cells survived and participated in capillary networks in the ischemic tissues of immunodeficient nude rats in vivo. AT cells thus had multiple endothelial phenotypes and were defined as a major population of EPCs. Furthermore, laser Doppler and immunohistochemical analyses revealed that EPC transplantation quantitatively augmented neovascularization and blood flow in the ischemic hindlimb. In conclusion, umbilical cord blood is a precious source for isolating EPCs, and transplantation of cord blood-derived EPCs would be a novel strategy to modulate postnatal neovascularization.

内皮前体细胞（EPCs）在成人外周血中已被发现。我们研究了内皮祖细胞是否可以从脐带血（造血祖细胞的丰富来源）中分离，以及内皮祖细胞的体内移植是否可以调节出生后的新生血管形成。脐带血单个核细胞（MNCs）培养后可形成大量细胞簇、梭形和贴壁（AT）细胞以及索状结构。示踪实验显示，细胞簇、AT细胞和索状结构主要来源于CD 34阳性MNC（MNC <CD 34 +>）。更多数量的AT细胞和细胞簇从脐带血-MNCs比从等量的成人外周血-MNCs。AT细胞掺入乙酰化LDL，释放一氧化氮，并表达KDR，VE-钙粘蛋白，CD 31和血管性血友病因子，但不表达CD 45。局部移植的AT细胞在免疫缺陷裸鼠缺血组织中存活并参与毛细血管网络。因此，AT细胞具有多种内皮表型，并被定义为EPCs的主要群体。此外，激光多普勒和免疫组化分析显示，EPC移植定量增加缺血后肢的新血管形成和血流。总之，脐带血是分离内皮祖细胞的宝贵来源，脐带血来源的内皮祖细胞的移植将是一种新的策略，以调节出生后的新生血管。

项目成果

Urano H, Ikeda H, Ueno T, Matsumoto T, Murohara T, Imaizumi T.: "Enhanced external counterpulsation improves exercise tolerance, reduces exercise-induced myocardial ischemia and improves left ventricular diastolic filling in patients with coronary artery

Urano H、Ikeda H、Ueno T、Matsumoto T、Murohara T、Imaizumi T.：“增强体外反搏可改善冠状动脉患者的运动耐量、减少运动引起的心肌缺血并改善左心室舒张期充盈

Murohara T, Ikeda H, Duan J, Shintani S, Sasaki Ki, Eguchi H, Onitsuka I, Matsui K, Imaizumi T.: "Transplanted cord blood-derived endothelial precursor cells augment postnatal neovascularization."J Clin Invest. 105. 1527-1536 (2000)

Murohara T、Ikeda H、Duan J、Shintani S、Sasaki Ki、Eguchi H、Onitsuka I、Matsui K、Imaizumi T.：“移植的脐带血来源的内皮前体细胞增强了产后新生血管形成。”J Clin Invest。

Ikeda H,Ueyama T,Murohara T,et al.: "Adhesive interaction between P-selectin and sialyl lewisX plays an important role in recurrent coronary arterial thrombosis in dogs."Arterioscler.Thromb.Vasc.Biol.. 19. 1083-1090 (1999)

Ikeda H、Ueyama T、Murohara T 等人：“P-选择素和唾液酸 lewisX 之间的粘附相互作用在狗的复发性冠状动脉血栓形成中发挥着重要作用。”Arterioscler.Thromb.Vasc.Biol.. 19. 1083-1090

Duan J, Murohara T, Ikeda H, Sasaki Ki, Shintani S, Akita T, Shimada T, Imaizumi T.: "Hyperhomocysteinemia impairs angiogenesis in response to hindlimb ischemia."Arterioscler Thromb Vasc Biol. 20. 2579-2585 (2000)

Duan J、Murohara T、Ikeda H、Sasaki Ki、Shintani S、Akita T、Shimada T、Imaizumi T.：“高同型半胱氨酸血症会损害后肢缺血反应中的血管生成。”动脉硬化血栓血管生物学。

Takajo Y, Ikeda H, Haramaki N, Murohara T, Imaizumi T.: "Augmented oxidative stress of platelets in chronic smokers. Mechanisms of impaired platelet-derived nitric oxide bioactivity and augmented platelet aggregability."J Am Coll Cardiol. (in press). (200

Takajo Y、Ikeda H、Haramaki N、Murohara T、Imaizumi T.：“慢性吸烟者血小板氧化应激增强。血小板源性一氧化氮生物活性受损和血小板聚集性增强的机制。”J Am Coll Cardiol。