A protocol for phenotypic detection and enumeration of circulating endothelial cells and circulating progenitor cells in human blood
A protocol for phenotypic detection and enumeration of circulating endothelial cells and circulating progenitor cells in human blood
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DOI：
10.1038/nprot.2007.111
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Jain, Rakesh K.
中科院分区：
文献类型：
--
作者：
Duda, Dan G.;Cohen, Kenneth S.;Jain, Rakesh K.
血液循环内皮细胞（CEC）和循环造血祖细胞（CPC）代表了被认为在组织血管化中起重要作用的两种细胞群体。CEC和CPC目前被研究作为十几种病理学的替代标志物，包括心脏病和癌症。然而，由于不同实验室用于评估和计数这些细胞的多种定义、方法和协议，数据解释通常是困难的。在这里，我们提出了一个细胞仪的协议，用于表型鉴定和计数CEC和CPC在人类血液中使用四个表面标记：CD 31，CD 34，CD 133和CD 45。这种方法允许进一步的表型分析，以探索这些细胞的生物学。此外，它还为不同病理患者的这些细胞的纵向研究提供了平台。该方案相对简单、便宜，并且可以适用于多种流式细胞仪类型或软件。该程序应持续2-2.5小时，预计可检测血液单核细胞群中0.1-6.0%的活CEC和0.01-0.20%的CPC。
Blood circulating endothelial cells (CECs) and circulating hematopoietic progenitor cells (CPCs) represent two cell populations that are thought to play important roles in tissue vascularization. CECs and CPCs are currently studied as surrogate markers in patients for more than a dozen pathologies, including heart disease and cancer. However, data interpretation has often been difficult because of multiple definitions, methods and protocols used to evaluate and count these cells by different laboratories. Here, we propose a cytometry protocol for phenotypic identification and enumeration of CECs and CPCs in human blood using four surface markers: CD31, CD34, CD133 and CD45. This method allows further phenotypic analyses to explore the biology of these cells. In addition, it offers a platform for longitudinal studies of these cells in patients with different pathologies. The protocol is relatively simple, inexpensive and can be adapted for multiple flow cytometer types or software. The procedure should take 2-2.5 h, and is expected to detect 0.1-6.0% viable CECs and 0.01-0.20% CPCs within blood mononuclear cell population.