López-Holgado N, Alberca M, Sánchez-Guijo F, Villarón E, Almeida J, Martín A, Armellini A, García C, Blanco B, Sánchez-Abarca I, Martín S, Pérez-Simón J A, García-Sanz R, San Miguel J F, del Cañizo M C
Department of Hematology, Hospital Clínico Universitario de Salamanca, Salamanca, Spain.
Cytotherapy. 2007;9(1):14-22. doi: 10.1080/14653240601047726.
The aim of this study was to identify circulating endothelial progenitor cells (EPC) with colony-forming capacity and compare them with the monocytic-macrophage lineage.
Forty-two healthy donors were analyzed. EPC were cultured with VEGF and b-FGF. Sequential studies were performed on days +7 (colonies) +21 and +35. Monocytic cells were cultured using the same conditions as EPC until day +21 or alternatively by adding IGF.
The number of EPC colonies was higher in BM than in mobilized or steady-state PB. Using EPC medium, monocytic cells formed cord-like structures but no colonies. However, colonies grew when IGF was added to the medium. By immunocytochemistry, colonies showed CD45, CD31 and lysozyme but no vWF. Colonies were CD4+, CD13+dim, CD14+, CD15++, CD16-/+dim, CD31+dim, CD33+dim, CD45+, CD105-/+dim, lysozyme+ and VE-cadherin+, and constantly negative for CD34, CD133 and KDR, when flow cytometry was used. The immunophenotype of pre-cultured and cultured monocytes was similar to that described for EPC.
Our results suggest that the so-called 'EPC' obtained at 7 days of culture belong to the monocyte-macrophage lineage, as they share immunophenotypic and molecular features.
本研究旨在鉴定具有集落形成能力的循环内皮祖细胞（EPC），并将其与单核巨噬细胞谱系进行比较。
分析了42名健康供体。EPC用血管内皮生长因子（VEGF）和成纤维细胞生长因子（b-FGF）培养。在第7天（集落）、第21天和第35天进行序贯研究。单核细胞采用与EPC相同的条件培养至第21天，或者添加胰岛素样生长因子（IGF）进行培养。
骨髓中EPC集落的数量高于动员或稳态外周血中的数量。使用EPC培养基时，单核细胞形成索状结构但无集落。然而，向培养基中添加IGF后集落生长。通过免疫细胞化学检测，集落显示CD45、CD31和溶菌酶阳性，但血管性血友病因子（vWF）阴性。流式细胞术检测时，集落为CD4+、CD13+弱阳性、CD14+、CD15++、CD16-/+弱阳性、CD31+弱阳性、CD33+弱阳性、CD45+、CD105-/+弱阳性、溶菌酶+和血管内皮钙黏蛋白（VE-cadherin）+，且持续CD34、CD133和激酶插入结构域受体（KDR）阴性。预培养和培养后的单核细胞免疫表型与EPC相似。
我们的结果表明，培养7天时获得的所谓“EPC”属于单核巨噬细胞谱系，因为它们具有共同的免疫表型和分子特征。
