Effect of flow and surface conditions on human lymphocyte isolation using microfluidic chambers
Effect of flow and surface conditions on human lymphocyte isolation using microfluidic chambers
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DOI：
10.1021/la048047b
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发表时间：
2004-12-21
期刊：
影响因子：
3.9
通讯作者：
Toner, M
中科院分区：
文献类型：
--
作者：
Murthy, SK;Sin, A;Toner, M
表型纯净的淋巴细胞亚群可以提供对损伤和疾病的免疫反应的有价值的见解。这些亚群的分离带来了独特的挑战，特别是当要最大限度地减少标记荧光或抗体的前处理孵化时。本文研究了使用涂有抗体的微流体室从混合物中分离T和B淋巴细胞，重点研究了流动条件和表面化学。两种细胞的粘附力随着剪切力的增加而减少，而与表面化学无关。聚乙二醇链的加入以及小室表面的抗体被证明显著改善了细胞黏附的重复性，因此是整个系统设计的重要部分。此外，该技术被证明是一种从模型混合物中分离高纯度淋巴细胞亚群的有效方法，即使在目标细胞浓度较低的情况下也是如此。
Phenotypically pure subpopulations of lymphocytes can provide valuable insights into the immune response to injury and disease. The isolation of these subpopulations presents unique challenges, particularly when preprocessing incubation to attach fluorescent or antibody tags is to be minimized. This paper examines the separation of T and B lymphocytes from mixtures using microfluidic chambers coated with antibodies, focusing on flow conditions and surface chemistry. The adhesion of both cell types decreases as shear stress increases irrespective of the surface chemistry. The incorporation of poly(ethylene glycol) chains along with the antibodies on the chamber surface is shown to significantly improve the reproducibility of cell adhesion and is thus an important part of the overall system design. Furthermore, this technique is shown to be an effective way of isolating highly pure subpopulations of lymphocytes from model mixtures, even when the target cell concentration is low.