Microfluidic isolation of leukocytes from whole blood for phenotype and gene expression analysis
Microfluidic isolation of leukocytes from whole blood for phenotype and gene expression analysis
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DOI：
10.1021/ac060140c
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发表时间：
2006-08-01
影响因子：
7.4
通讯作者：
Toner, Mehmet
中科院分区：
文献类型：
--
作者：
Sethu, Palaniappan;Moldawer, Lyle L.;Toner, Mehmet
能够从复杂群体的小样本中分离细胞亚型的技术将极大地促进蛋白质组学和基因组学在人类疾病中的应用。血液转录组分析需要去除污染红细胞。我们报告了一种自动化微流体装置，可通过去离子水裂解快速消耗全血中的红细胞，并收集富集的白细胞用于表型和基因组分析。从健康受试者的血液开始，我们展示了这种微流体盒和裂解方案在制备未刺激的白细胞以及用葡萄球菌肠毒素 B 离体刺激的白细胞方面的实用性，它模仿了严重细菌感染患者中观察到的一些细胞效应。微阵列用于评估对肠毒素 B 的整体基因表达反应。结果表明，该系统可以从少量血液样本中分离出未激活的白细胞，而不会造成任何显着损失，这使得可以从后续分析中获得更多信息，并且很容易应用于临床环境。
Technologies that enable the isolation of cell subtypes from small samples of complex populations will greatly facilitate the implementation of proteomics and genomics to human diseases. Transcriptome analysis of blood requires the depletion of contaminating erythrocytes. We report an automated microfluidic device to rapidly deplete erythrocytes from whole blood via deionized water lysis and to collect enriched leukocytes for phenotype and genomic analyses. Starting with blood from healthy subjects, we demonstrate the utility of this microfluidic cassette and lysis protocol to prepare unstimulated leukocytes, and leukocytes stimulated ex vivo with Staphylococcal enterotoxin B, which mimics some of the cellular effects seen in patients with severe bacterial infections. Microarrays are used to assess the global gene expression response to enterotoxin B. The results demonstrate that this system can isolate unactivated leukocytes from small blood samples without any significant loss, which permits more information to be obtained from subsequent analysis, and will be readily applicable to clinical settings.