Cytometry：EdU检测细胞增殖效果最佳

　　

北京协和医院研究者利用EdU检测试剂结合流式细胞分析来检测T-淋巴细胞在体外的增殖情况，并优化出最佳的测定条件。研究者这一系统的研究为今后的细胞增殖的检测提供了重要的实验依据，相关研究发表在Cytometry杂志。

EdU是一种胸腺嘧啶核苷类似物，能够在DNA复制时期代替胸腺嘧啶(T)渗入正在合成的DNA分子中，通过基于荧光染料与EdU的特异性反应即可直接检测出DNA复制活性，是一种新型的细胞增殖检测方法。与传统的（3H）胸腺嘧啶检测及BrdU检测方法相比，EdU检测方法在体内、体外的细胞动力学、DNA合成及细胞增殖检测中均表现出很大的优势。

研究者采用锐博生物EdU细胞增殖检测试剂（Cell-Light™ EdU Cell Proliferation Detection），经过一系列严密的实验研究，发现细胞增殖检测效果与EdU浓度、孵化时间、试剂体积有很大关联。最终获得检测T-淋巴细胞增殖EdU试剂的最佳浓度为10-50 μM，最佳孵化时间为8-12h，最佳反应体积为100 μl。采用含0.05% 皂素的PBS溶液去除细胞通透性试剂的效果更好。加入EdU试剂的T-淋巴细胞可在液氮状态下可保存长达21天。这一系列研究为采用EdU结合流式细胞仪检测细胞增殖提供了最佳检测方案，为研究T-淋巴细胞增殖及标记细胞表面抗原等实验开展有重要指导意义。

锐博生物EdU细胞增殖检测试剂更简单、更灵敏、更快速、更准确，是细胞增殖检测的最佳选择。此外，锐博生物经验丰富的核酸研发团队还提供一系列miRNA产品及服务，包括，miRNA mimic、miRNA inhibitor、miRNA agomir、miRNA antagomir、miRNA qRT-PCR、miRNA表达载体构建、miRNA靶基因预测分析、miRNA表达谱芯片、miRNA新一代高通量测序等，为研究者提供全方位的技术服务支持及完善的实验解决方案。

锐博生物推荐原文摘要：

Multicolor Flow Cytometry Analysis of the Proliferations of T-Lymphocyte Subsets In Vitro by EdU Incorporation

Cytometry A. 2012 Oct;81(10):901-9. Epub 2012 Aug 28.

Abstract：EdU (5-ethynyl-2'-deoxyuridine) incorporation has proved advantageous in the studies of cell kinetics, DNA synthesis, and cellular proliferation in vitro and in vivo compared to [(3) H]thymidine incorporation and BrdU (5-bromo-2'-deoxyuridine) incorporation. Here, we describe a method that combines EdU incorporation and immunostaining with flow cytometric analysis to detect the proliferations of T lymphocyte subsets in vitro and optimized the assay's conditions. We found that the number of EdU(+) cells were associated with EdU concentration, incubation time, and the volume of Click reaction solution, the best EdU concentration 10-50 μM, the optimal incubation time 8-12 h and the proper volume of Click volume 100 μl for labeling 1 × 10(6) lymphocytes. Fixation was better to be performed before permeabilization, not together with. Furthermore, the permeabilization detergent reagent, PBS with 0.05% saponin was better than Tris buffer saline (TBS) with 0.1% Triton X-100. In addition, sufficient wash with PBS with 0.05% saponin has no influence on the staining of EdU(+) cells. Also, the lymphocytes incorporating EdU could be stored at 4°C, -80°C, and in liquid nitrogen up to 21 days. The present study will aid in optimization of flow cytometry assay to detect the proliferations of T cell subsets by EdU incorporation and the labeling of cell surface antigens.

http://www.ncbi.nlm.nih.gov/pubmed?term=Multicolor%20Flow%20Cytometry%20Analysis%20of%20the%20Proliferations%20of%20T-Lymphocyte%20Subsets%20In%20Vitro%20by%20EdU%20Incorporation