Redondo Santiago, Hristov Mihail, Gordillo-Moscoso Antonio A, Ruiz Emilio, Weber Christian, Tejerina Teresa
Department of Pharmacology, School of Medicine, Universidad Complutense, Madrid, Spain.
J Immunol Methods. 2008 Jun 1;335(1-2):21-7. doi: 10.1016/j.jim.2008.02.011. Epub 2008 Mar 19.
Although determination of circulating endothelial progenitor cell (EPC) in peripheral blood by flow cytometry is an emerging marker for cardiovascular medicine, a common standardized protocol is still not available, due to the low numbers achieved in peripheral blood. In the present paper we describe a novel technique for EPC quantification as CD34+/CD144+/CD3- cells within the lymphocyte gate, which increases the percentages of EPC positivity described before and also offers high intra-assay reproducibility. These improvements are based on a gating strategy for big-sized lymphocytes, smooth fixation and cytometric clearance of CD3+ lymphocytes (T-cells). This last procedure is able to increase intra-assay Pearson's correlation from 0.8517 to 0.8908. Therefore, the technical setting described here offers a high-performance and clinically oriented EPC determination strategy in human peripheral blood.
尽管通过流式细胞术测定外周血中循环内皮祖细胞（EPC）是心血管医学中一种新兴的标志物，但由于外周血中EPC数量较少，目前仍没有通用的标准化方案。在本文中，我们描述了一种新的技术，用于将EPC定量为淋巴细胞门内的CD34+/CD144+/CD3-细胞，该技术提高了之前所描述的EPC阳性百分比，并且具有较高的批内重复性。这些改进基于对大尺寸淋巴细胞的设门策略、平滑固定以及对CD3+淋巴细胞（T细胞）的细胞计数清除。最后这一步骤能够将批内Pearson相关性从0.8517提高到0.8908。因此，本文所描述的技术设置为人类外周血中EPC的测定提供了一种高性能且面向临床的策略。
