DNAse treatment following thawing of Cryopreserved PBMC is a procedure suitable for lymphocyte functional studies
DNAse treatment following thawing of Cryopreserved PBMC is a procedure suitable for lymphocyte functional studies
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DOI：
10.1016/j.jim.2006.04.004
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发表时间：
2006-06-30
影响因子：
2.2
通讯作者：
Pinto, Ligia A.
中科院分区：
文献类型：
--
作者：
Garcia-Pineres, Alfonso J.;Hildesheim, Allan;Pinto, Ligia A.
在大型临床试验或自然病史研究中，检测新分离的PBMC不适用于免疫监测分析。因此，深低温保存的PBMC是一种更实用的替代方法。然而，从以前运输和储存的血液中分离出来的PBMC解冻后，细胞聚集是一个常见的问题。细胞聚集会导致细胞丢失，并可能影响细胞功能和/或表型。为了评估临床研究中冷冻保存的样本的免疫功能和表型，有必要开发和验证防止细胞聚集并保持细胞功能和表面标记表达水平的程序。在标准解冻程序中加入DNA酶处理步骤有效地避免了集落的形成，并且不会导致细胞活力、标准白细胞表面标记的表达或免疫功能、增殖和细胞因子诱导的两个关键参数对各种常见刺激的可检测到变化。因此，这一程序似乎适用于标准的免疫学检测。(C)2006爱思唯尔B.V.保留所有权利。
Testing freshly isolated PBMC is not practical for immune monitoring analysis in large clinical trials or natural history studies. Thus, cryopreserved PBMC represent a more practical alternative. However, cell clumping is a common problem following thawing of PBMC isolated from blood that was previously transported and stored. Cell clumping leads to loss of cells, and could affect cell function and/or phenotype. The development and validation of procedures that prevent cell clumping and preserve cell function and surface marker expression levels are necessary to allow evaluation of immune function and phenotype in cryopreserved samples from clinical studies.The incorporation of a DNAse treatment step in the standard thawing procedure efficiently avoided clump formation, and did not result in detectable changes in cell viability, expression of standard leukocyte surface markers or two key parameters of immune function, proliferation and cytokine induction in response to a variety of common stimuli. Therefore, this procedure seems suitable for standard immunologic assays. (c) 2006 Elsevier B.V. All rights reserved.