Effects of EdU labeling on mesenchymal stem cells.
Effects of EdU labeling on mesenchymal stem cells.
复制标题
DOI：
10.1016/j.jcyt.2012.10.010
复制
发表时间：
2013-01
期刊：
影响因子：
4.5
通讯作者：
Lin, Ching-Shwun
中科院分区：
文献类型：
--
作者：
Ning, Hongxiu;Albersen, Maarten;Lin, Guiting;Lue, Tom F.;Lin, Ching-Shwun
关键词：
胸苷类似物 5-乙炔基-2-脱氧尿苷 (EdU) 最近被用于追踪间充质干细胞 (MSC)。在本研究中，我们测试了 EdU 是否具有细胞毒性，以及它是否干扰 MSC 的分化、细胞因子分泌和迁移。 EdU 标记是通过将脂肪干细胞 (ADSC) 与 10 μM EdU 一起孵育 48 小时来进行的。通过与叠氮化物缀合的 Alexa594 反应检测 EdU 的掺入。比较标记和未标记的 ADSC 的增殖和凋亡，分别通过 CellTiter 和彗星测定法测定。还根据形态、标记表达和功能对它们进行了神经元样和内皮分化的比较。通过细胞因子抗体阵列对它们分泌的细胞因子谱进行比较。通过迁移测定比较它们对归巢因子 SDF-1 的反应。大约 70% 的 ADSC 中 EdU 被整合到细胞核中。 EdU 标记和未标记的 ADSC 之间没有观察到增殖和凋亡率的显着差异。异丁基甲基黄嘌呤 (IBMX) 诱导 EdU 标记和未标记的 ADSC 呈现神经元样形态并表达 β-III 微管蛋白。内皮生长培养基 2 (EGM2) 诱导 EdU 标记和未标记的 ADSC 中的内皮分化，包括摄取低密度脂蛋白 (LDL) 和形成毛细血管样结构的能力以及 vWF、eNOS 和 CD31 的表达。 EdU 标记和未标记的 ADSC 表现出相同的分泌细胞因子谱和对 SDF-1 的相同迁移反应。推荐剂量 10 μM EdU 对 ADSC 无毒。 EdU 标签不会干扰 ADSC 的分化、细胞因子分泌或对 SDF-1 的迁移反应。
Thymidine analog 5-ethynyl-2-deoxyuridine (EdU) has recently been employed for tracking mesenchymal stem cells (MSCs). In the present study we tested whether EdU was cytotoxic and whether it interfered with MSC’s differentiation, cytokine secretion, and migration. EdU labeling was performed by incubating adipose-derived stem cells (ADSCs) with 10 μM of EdU for 48 hours. Incorporation of EdU was detected by reaction with azide-conjugated Alexa594. The labeled and unlabeled ADSCs were compared for proliferation and apoptosis as determined by CellTiter and comet assays, respectively. They were also compared for neuron-like and endothelial differentiation as determined by morphology, marker expression, and function. Comparison of their secreted cytokine profile was performed by cytokine antibody array. Comparison of their response to homing factor SDF-1 was performed by migration assay. EdU was incorporated into the nucleus in approximately 70% of ADSCs. No significant differences in proliferation and apoptosis rates were observed between EdU-labeled and unlabeled ADSCs. Isobutylmethylxanthine (IBMX) induced both EdU-labeled and unlabeled ADSCs to assume a neuron-like morphology and to express β-III tubulin. Endothelial growth medium-2 (EGM2) induced endothelial differentiation in both EdU-labeled and unlabeled ADSCs, including the ability to uptake low-density lipoprotein (LDL) and to form capillary-like structures as well as the expression of vWF, eNOS, and CD31. EdU-labeled and unlabeled ADSCs exhibited identical secreted cytokine profile and identical migratory response to SDF-1. At the recommended dosage of 10 μM EdU is non-toxic to ADSCs. EdU label did not interfere with ADSC’s differentiation, cytokine secretion, or migratory response to SDF-1.
登录
查看更多内容
影响因子：
4.3
作者：
Lequeux, Charlotte;Oni, Georgette;Brown, Spencer A.
通讯作者：
Brown, Spencer A.
DOI：
10.1111/j.1743-6109.2009.01697.x
发表时间：
2010-04
期刊：
The journal of sexual medicine
影响因子：
--
作者：
Huang YC;Ning H;Shindel AW;Fandel TM;Lin G;Harraz AM;Lue TF;Lin CS
通讯作者：
Lin CS
DOI：
10.1016/j.bbrc.2010.10.090
发表时间：
2010-11-19
影响因子：
3.1
作者：
Zhang H;Ning H;Banie L;Wang G;Lin G;Lue TF;Lin CS
通讯作者：
Lin CS
影响因子：
5.3
作者：
Xu J;Liu X;Jiang Y;Chu L;Hao H;Liua Z;Verfaillie C;Zweier J;Gupta K;Liu Z
通讯作者：
Liu Z
影响因子：
2.9
作者：
Ning, Hongxiu;Lin, Guiting;Lin, Ching-Shwun
通讯作者：
Lin, Ching-Shwun