Isolation of whole mononuclear cells from peripheral blood and cord blood.
Isolation of whole mononuclear cells from peripheral blood and cord blood.
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DOI：
10.1002/0471142735.im0701s85
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发表时间：
2009-04-01
影响因子：
--
通讯作者：
Zola, Heddy
中科院分区：
文献类型：
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作者：
Fuss, Ivan J;Kanof, Marjorie E;Zola, Heddy
外周血是研究人类免疫系统的淋巴细胞的主要来源。Ficoll-Hypaque密度梯度离心法是一种简单快速的纯化外周血单核细胞（PBMC）的方法，利用了血液样本中单核细胞和其他元素之间的密度差异。因此，细胞基于其密度/尺寸的差异分层分布在溶液中。可以采用其他纯化方法，因为单核细胞样品可以通过粘附或通过暴露于L-亮氨酸甲酯从单核细胞中纯化;这些方法描述了两种程序。来自婴儿的脐带血和外周血含有未成熟细胞，包括有核红细胞，这可能导致单核细胞层的显著污染，并且去除这些细胞需要描述的额外步骤。本文提出的分离方法也可应用于来源于组织的细胞群。
Peripheral blood is the primary source of lymphoid cells for investigation of the human immune system. Its use is facilitated by Ficoll-Hypaque density gradient centrifugation-a simple and rapid method of purifying peripheral blood mononuclear cells (PBMC) that takes advantage of the density differences between mononuclear cells and other elements found in the blood sample. Thus, cells are distributed in the solution in layers based on the differences in their density/size. Additional purification methods can be employed as the mononuclear cell sample can be purified from monocytes by adherence or by exposure to L-leucine methyl ester; these methods are described for both procedures. Cord blood and peripheral blood from infants contain immature cells, including nucleated red cells, which can result in significant contamination of the mononuclear cell layer, and removal of these cells requires additional steps that are described. The isolation procedures presented here can also be applied to cell populations derived from tissues.