Human cord blood-derived AC133+ progenitor cells preserve endothelial progenitor characteristics after long term in vitro expansion.
Human cord blood-derived AC133+ progenitor cells preserve endothelial progenitor characteristics after long term in vitro expansion.
复制标题
DOI：
10.1371/journal.pone.0009173
复制
发表时间：
2010-02-11
期刊：
影响因子：
3.7
通讯作者：
Arbab AS
中科院分区：
文献类型：
--
作者：
Janic B;Guo AM;Iskander AS;Varma NR;Scicli AG;Arbab AS
干细胞/祖细胞是血管缺血性疾病细胞治疗方法发展的核心。从缺血中拯救组织的关键步骤是通过促进新血管形成来改善血管形成。内皮祖细胞（EPCs）是开发这种方法的最佳候选者，因为它们具有自我更新、循环和分化为成熟内皮细胞（ECs）的能力。研究表明，静脉注射从骨髓、外周血或脐带血中分离的祖细胞可返回缺血部位。然而，这种移植疗法的成功临床应用受到可以从患者产生的少量EPCs的限制。因此，通过长期体外扩增扩增自体EPC的数量同时保持其血管生成潜力的能力对于开发基于EPC的疗法至关重要。因此，本研究的目的是评估脐带血（CB）来源的AC 133+细胞在体外长期扩增后向功能性成熟内皮细胞（EC）分化的能力。我们系统地表征了CB AC 133+细胞在体外扩增30天内的特性。在30天的培养过程中，CB AC 133+细胞表现出显著的生长潜力，表现为细胞数量增加了148倍。流式细胞术和免疫细胞化学结果显示，长期体外培养对CB AC 133+细胞内皮祖细胞标志物的表达无影响。在EC分化条件下培养后，细胞表现出成熟EC标志物的高表达，如CD 31、VEGFR-2和von Willebrand因子，以及指示向成熟EC分化的形态学变化。此外，在整个30天的培养细胞保留其功能的能力，证明了高摄取的DiI荧光共轭乙酰化低密度脂蛋白（DiI-Ac-LDL），在体外和体内迁移趋化刺激和体外管形成。这些研究表明，原代CB AC 133+培养物主要含有EPC，并且长期体外条件有利于维持这些细胞处于向内皮谱系定型的状态。
Stem cells/progenitors are central to the development of cell therapy approaches for vascular ischemic diseases. The crucial step in rescuing tissues from ischemia is improvement of vascularization that can be achieved by promoting neovascularization. Endothelial progenitor cells (EPCs) are the best candidates for developing such an approach due to their ability to self-renew, circulate and differentiate into mature endothelial cells (ECs). Studies showed that intravenously administered progenitors isolated from bone marrow, peripheral or cord blood home to ischemic sites. However, the successful clinical application of such transplantation therapy is limited by low quantities of EPCs that can be generated from patients. Hence, the ability to amplify the numbers of autologous EPCs by long term in vitro expansion while preserving their angiogenic potential is critically important for developing EPC based therapies. Therefore, the objective of this study was to evaluate the capacity of cord blood (CB)-derived AC133+ cells to differentiate, in vitro, towards functional, mature endothelial cells (ECs) after long term in vitro expansion. We systematically characterized the properties of CB AC133+ cells over the 30 days of in vitro expansion. During 30 days of culturing, CB AC133+ cells exhibited significant growth potential that was manifested as 148-fold increase in cell numbers. Flow cytometry and immunocytochemistry demonstrated that CB AC133+ cells' expression of endothelial progenitor markers was not affected by long term in vitro culturing. After culturing under EC differentiation conditions, cells exhibited high expression of mature ECs markers, such as CD31, VEGFR-2 and von Willebrand factor, as well as the morphological changes indicative of differentiation towards mature ECs. In addition, throughout the 30 day culture cells preserved their functional capacity that was demonstrated by high uptake of DiI fluorescently conjugated-acetylated-low density lipoprotein (DiI-Ac-LDL), in vitro and in vivo migration towards chemotactic stimuli and in vitro tube formation. These studies demonstrate that primary CB AC133+ culture contained mainly EPCs and that long term in vitro conditions facilitated the maintenance of these cells in the state of commitment towards endothelial lineage.
登录
查看更多内容
影响因子：
56.9
作者：
Asahara, T;Murohara, T;Isner, JM
通讯作者：
Isner, JM
DOI：
10.1007/bf01476669
发表时间：
1978-01-01
期刊：
KLINISCHE WOCHENSCHRIFT
影响因子：
--
作者：
HLADOVEC, J;PREROVSKY, I;FABIAN, J
通讯作者：
FABIAN, J
影响因子：
20.1
作者：
Fujiyama, S;Amano, K;Matsubara, H
通讯作者：
Matsubara, H
影响因子：
82.9
作者：
Ceradini, DJ;Kulkarni, AR;Gurtner, GC
通讯作者：
Gurtner, GC
DOI：
10.1084/jem.20040440
发表时间：
2004-07-19
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Kögler G;Sensken S;Airey JA;Trapp T;Müschen M;Feldhahn N;Liedtke S;Sorg RV;Fischer J;Rosenbaum C;Greschat S;Knipper A;Bender J;Degistirici O;Gao J;Caplan AI;Colletti EJ;Almeida-Porada G;Müller HW;Zanjani E;Wernet P
通讯作者：
Wernet P