Retention of a differentiated endothelial phenotype by outgrowth endothelial cells isolated from human peripheral blood and expanded in long-term cultures
Retention of a differentiated endothelial phenotype by outgrowth endothelial cells isolated from human peripheral blood and expanded in long-term cultures
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DOI：
10.1007/s00441-006-0222-4
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发表时间：
2006-10-01
影响因子：
3.6
通讯作者：
Kirkpatrick, Charles James
中科院分区：
文献类型：
--
作者：
Fuchs, Sabine;Hermanns, Maria Iris;Kirkpatrick, Charles James
自体人内皮细胞的快速充分血管化仍然是治疗缺血组织和生成新组织的限制步骤。我们从人外周血中扩增了生长内皮细胞 (OEC)，并研究了它们在长期培养中的表型稳定性。我们的目标是获得合适数量的自体内皮细胞用于促血管生成细胞疗法。从人外周血中分离出单核细胞。在培养过程中，在与成熟内皮细胞的单培养或共培养中对细胞进行了多种内皮细胞和干细胞标记物的表征。在外周血培养物中，我们观察到细胞具有不同的呈现分化内皮表型的能力。大多数细胞显示出内皮祖细胞或成血管细胞的标记物（CD31、KDR、VE-钙粘蛋白、CD34、CD117、CD45），但未能形成分化表型。通过逆转录/聚合酶链反应 (RT-PCR) 或免疫荧光在这些细胞中未检测到 Caveolin-1。另一种由相同培养物产生的细胞类型表达分化表型，被命名为 OEC。该子集作为 OEC 在长期培养中得到扩展，并通过免疫荧光、流式细胞术和 RT-PCR 进行分析以获得稳定的内皮表型。 OEC 显示出多种分化内皮细胞的标志物，例如所有测试传代中的 Caveolin-1 水平较高，以及在体外形成血管生成芽的能力。因此，来自血液单核细胞的长期扩增培养物中的OEC表型高度稳定，这一特征是使用来自外周血的OEC进行自体内皮细胞治疗的重要先决条件。
Rapid adequate vascularization by autologous human endothelial cells remains a limiting step in the treatment of ischemic tissues and the generation of new tissues. We have expanded outgrowth endothelial cells (OEC) from human peripheral blood and investigated their phenotypic stability in long-term cultures. Our goal has been to obtain suitable numbers of autologous endothelial cells for pro-angiogenic cell therapies. Mononuclear cells were isolated from human peripheral blood. During culture, cells were characterized for several endothelial and stem cell markers in mono- or in co-culture with mature endothelial cells. In cultures from peripheral blood, we observed cells with a variable ability to assume a differentiated endothelial phenotype. Most of the cells showed markers reported for endothelial progenitor cells or hemangioblasts (CD31, KDR, VE-cadherin, CD34, CD117, CD45) but failed to develop a differentiated phenotype. Caveolin-1 was not detectable in these cells by reverse transcription/polymerase chain reaction (RT-PCR) or immunofluorescence. Another cell type arising from the same cultures expressed a differentiated phenotype and was designated as an OEC. This subset as an OEC was expanded in long-term cultures and analyzed by immunofluorescence, flow-cytometry, and RT-PCR for a stable endothelial phenotype. OEC showed several markers of a differentiated endothelium, such as high levels of caveolin-1 throughout all tested passages, and the ability to form angiogenic sprouts in vitro. Thus, OEC in long-term expansion cultures from blood mononuclear cells are phenotypically highly stable, a feature that is an important prerequisite for using OEC from peripheral blood for autologous endothelial cell therapies.