Phenotypic and functional characterization of endothelial colony forming cells derived from human umbilical cord blood.
Phenotypic and functional characterization of endothelial colony forming cells derived from human umbilical cord blood.
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DOI：
10.3791/3872
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发表时间：
2012-04
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影响因子：
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通讯作者：
Nutan Prasain;J. L. Meador;Mervin C Yoder
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文献类型：
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作者：
Nutan Prasain;J. L. Meador;Mervin C Yoder
长期以来关于血管生成、血管发生和动脉发生的新血管形成的观点最近得到了回顾。1997年，Asahara等人首次在成人外周血中发现了循环内皮祖细胞（EPCs）的存在，为血管再生和修复提供了新的假设和策略。内皮祖细胞是循环血液中罕见但正常的成分，它们位于血管形成或血管重塑的部位，并主要通过旁分泌刺激现有血管壁源性细胞促进出生后血管发生、血管生成或动脉发生。目前还没有发现特异性标记物来识别EPC，目前该领域的现状是了解许多细胞类型，包括促血管生成造血干细胞和祖细胞、循环血管生成细胞、Tie2+单核细胞、髓系祖细胞、肿瘤相关巨噬细胞和M2活化的巨噬细胞，在各种临床前动物模型系统和多种疾病状态的人类受试者中参与刺激血管生成过程。内皮集落形成细胞（Endothelial colony forming cells, ecfc）是一种罕见的循环存活内皮细胞，具有强大的克隆增殖潜力，在复制时具有二级和三级集落形成能力，并且在移植到免疫缺陷小鼠体内后能够形成体内固有血管。虽然ecfc已经成功地从健康成人的外周血、健康新生儿的脐带血（CB）和许多人类动脉和静脉血管的血管壁中分离出来。CB具有最高频率的ecfc，显示出最强大的克隆增殖潜力，并在体内形成持久和功能性的血管。虽然已经提出了从成人外周血中提取ECFC的方法，但在这里，我们描述了从人脐带CB中提取ECFC的方法，包括提取、克隆、扩增、体外和体内表征。
Longstanding views of new blood vessel formation via angiogenesis, vasculogenesis, and arteriogenesis have been recently reviewed. The presence of circulating endothelial progenitor cells (EPCs) were first identified in adult human peripheral blood by Asahara et al. in 1997 bringing an infusion of new hypotheses and strategies for vascular regeneration and repair. EPCs are rare but normal components of circulating blood that home to sites of blood vessel formation or vascular remodeling, and facilitate either postnatal vasculogenesis, angiogenesis, or arteriogenesis largely via paracrine stimulation of existing vessel wall derived cells. No specific marker to identify an EPC has been identified, and at present the state of the field is to understand that numerous cell types including proangiogenic hematopoietic stem and progenitor cells, circulating angiogenic cells, Tie2+ monocytes, myeloid progenitor cells, tumor associated macrophages, and M2 activated macrophages participate in stimulating the angiogenic process in a variety of preclinical animal model systems and in human subjects in numerous disease states. Endothelial colony forming cells (ECFCs) are rare circulating viable endothelial cells characterized by robust clonal proliferative potential, secondary and tertiary colony forming ability upon replating, and ability to form intrinsic in vivo vessels upon transplantation into immunodeficient mice. While ECFCs have been successfully isolated from the peripheral blood of healthy adult subjects, umbilical cord blood (CB) of healthy newborn infants, and vessel wall of numerous human arterial and venous vessels. CB possesses the highest frequency of ECFCs that display the most robust clonal proliferative potential and form durable and functional blood vessels in vivo. While the derivation of ECFC from adult peripheral blood has been presented, here we describe the methodologies for the derivation, cloning, expansion, and in vitro as well as in vivo characterization of ECFCs from the human umbilical CB.