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Functional comparison of PBMCs isolated by Cell Preparation Tubes (CPT) vs. Lymphoprep Tubes.
BMC Immunology ( IF 2.9 ) Pub Date : 2020-03-30 , DOI: 10.1186/s12865-020-00345-0
Han Chen 1 , Christian M Schürch 1 , Kevin Noble 2 , Kenneth Kim 3 , Peter O Krutzik 4 , Erika O'Donnell 4 , Jason Vander Tuig 4 , Garry P Nolan 1 , David R McIlwain 1, 2
Affiliation  

Cryopreserved human peripheral blood mononuclear cells (PBMCs) are a commonly used sample type for a variety of immunological assays. Many factors can affect the quality of PBMCs, and careful consideration and validation of an appropriate PBMC isolation and cryopreservation method is important for well-designed clinical studies. A major point of divergence in PBMC isolation protocols is the collection of blood, either directly into vacutainers pre-filled with density gradient medium or the use of conical tubes containing a porous barrier to separate the density gradient medium from blood. To address potential differences in sample outcome, we isolated, cryopreserved, and compared PBMCs using parallel protocols differing only in the use of one of two common tube types for isolation. Whole blood was processed in parallel using both Cell Preparation Tubes™ (CPT, BD Biosciences) and Lymphoprep™ Tubes (Axis-Shield) and assessed for yield and viability prior to cryopreservation. After thawing, samples were further examined by flow cytometry for cell yield, cell viability, frequency of 10 cell subsets, and capacity for stimulation-dependent CD4+ and CD8+ T cell intracellular cytokine production. No significant differences in cell recovery, viability, frequency of immune cell subsets, or T cell functionality between PBMC samples isolated using CPT or Lymphoprep tubes were identified. CPT and Lymphoprep tubes are effective and comparable methods for PBMC isolation for immunological studies.

中文翻译:

通过细胞制备管(CPT)与Lymphoprep管分离的PBMC的功能比较。

冷冻保存的人外周血单核细胞(PBMC)是用于各种免疫测定的常用样品类型。许多因素会影响PBMC的质量,对于精心设计的临床研究,仔细考虑和验证适当的PBMC分离和冷冻保存方法非常重要。PBMC隔离方案的主要区别点是血液收集,直接收集到预先装有密度梯度介质的真空容器中,还是使用包含多孔屏障的锥形管将密度梯度介质与血液分离。为了解决样品结果中的潜在差异,我们使用并行协议对PBMC进行了分离,冷冻保存和比较,其区别仅在于使用两种常见试管类型之一进行分离。使用Cell Preparation Tubes™(CPT,BD Biosciences)和Lymphoprep™管(Axis-Shield)并行处理全血,并在冷冻保存之前评估其产量和生存能力。解冻后,通过流式细胞术进一步检查样品的细胞产量,细胞活力,10个细胞亚群的频率以及刺激依赖性CD4 +和CD8 + T细胞内细胞因子产生的能力。在使用CPT或Lymphoprep管分离的PBMC样品之间,未发现细胞回收率,活力,免疫细胞亚群频率或T细胞功能方面的显着差异。CPT和Lymphoprep管是用于免疫学研究的PBMC分离的有效且可比的方法。通过流式细胞术进一步检查样品的细胞产量,细胞活力,10个细胞亚群的频率以及刺激依赖性CD4 +和CD8 + T细胞内细胞因子产生的能力。在使用CPT或Lymphoprep管分离的PBMC样品之间,未发现细胞回收率,活力,免疫细胞亚群频率或T细胞功能方面的显着差异。CPT和Lymphoprep管是用于免疫学研究的PBMC分离的有效且可比的方法。通过流式细胞术进一步检查样品的细胞产量,细胞活力,10个细胞亚群的频率以及刺激依赖性CD4 +和CD8 + T细胞内细胞因子产生的能力。在使用CPT或Lymphoprep管分离的PBMC样品之间,未发现细胞回收率,活力,免疫细胞亚群频率或T细胞功能方面的显着差异。CPT和Lymphoprep管是用于免疫学研究的PBMC分离的有效且可比的方法。鉴定了使用CPT或Lymphoprep管分离的PBMC样品之间的T细胞或T细胞功能。CPT和Lymphoprep管是用于免疫学研究的PBMC分离的有效且可比的方法。鉴定了使用CPT或Lymphoprep管分离的PBMC样品之间的T细胞或T细胞功能。CPT和Lymphoprep管是用于免疫学研究的PBMC分离的有效且可比的方法。
更新日期:2020-04-22
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