Biospecimen Science of Blood for Peripheral Blood Mononuclear Cell (PBMC) Functional Applications

被引:42
作者
Betsou, Fay [1 ]
Gaignaux, Amelie [1 ]
Ammerlaan, Wim [1 ]
Norris, Philip J. [2 ]
Stone, Mars [2 ]
机构
[1] IBBL, 1 Rue Louis Rech, L-3555 Dudelange, Luxembourg
[2] Vitalant Res Inst, 270 Masonic Ave, San Francisco, CA 94118 USA
关键词
PBMC; Human blood; Preanalytics; Functional assays; Cryopreservation; Quality control; POLYMERASE-CHAIN-REACTION; CRYOPRESERVATION IMPLICATIONS; LYMPHOCYTE IMMUNOPHENOTYPE; PROLIFERATIVE RESPONSES; QUALITY-ASSURANCE; INTERFERON-GAMMA; B-LYMPHOCYTES; WHOLE-BLOOD; ACTIVATION; STORAGE;
D O I
10.1007/s40139-019-00192-8
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
Purpose of Review Peripheral blood mononuclear cells (PBMCs) are used in a wide variety of preclinical assays. Preanalytical variations can have a devastating impact on the results. In this review, we list critical preanalytical factors for PBMC-based assays to develop awareness and orientation to the types of sample preparation and storage that one may consider employing. Recent Findings Critical factors during blood collection are the blood collection tube and anticoagulant, possible stabilizer used, and the pm-isolation blood storage temperature and time. During PBMC isolation, critical factors are the isolation method, density gradient or magnetic sorting, use of barrier, possible RBC lysis, and centrifuge type. During cryopreservation, attention is needed for the cryomedium type and temperature, freezing device and program, cell concentration, and the long-term storage temperature. During the thawing process, the thawing procedure/device used and wash medium temperature are critical. Summary To avoid biased results in PBMC assays, these critical preanalytical factors must be standardized and/or documented. Additionally, participation in external quality assurance programs is strongly recommended.
引用
收藏
页码:17 / 27
页数:11
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