Virtual Global Transplant Laboratory Standard Operating Procedures for Blood Collection, PBMC Isolation, and Storage
Virtual Global Transplant Laboratory Standard Operating Procedures for Blood Collection, PBMC Isolation, and Storage
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DOI：
10.1097/txd.0000000000000613
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发表时间：
2016-09-01
影响因子：
2.3
通讯作者：
Maltzman, Jonathan S.
中科院分区：
文献类型：
--
作者：
Higdon, Lauren E.;Lee, Karim;Maltzman, Jonathan S.
人类免疫反应的研究经常涉及在采集后立即或在明显延迟的时间点使用外周血单个核细胞（PBMC）。这需要从全血中分离PBMC，并在某些应用中进行低温保存。标准化血液采集、PBMC分离、低温保存和解冻的方案，在分析时最大限度地提高PBMC的存活率和功能是很重要的。该资源包括详细的协议描述采血管，分离PBMC使用密度梯度，PBMC的低温保存，和细胞的解冻以及准备功能分析。对于每个协议，我们包括重要的考虑因素，如时间，储存温度和冷冻速率。此外，我们提供了备选方案，以便研究人员可以在确定其应用程序的最佳协议时做出明智的决定。
Research on human immune responses frequently involves the use of peripheral blood mononuclear cells (PBMC) immediately, or at significantly delayed timepoints, after collection. This requires PBMC isolation from whole blood and cryopreservation for some applications. It is important to standardize protocols for blood collection, PBMC isolation, cryopreservation, and thawing that maximize survival and functionality of PBMC at the time of analysis. This resource includes detailed protocols describing blood collection tubes, isolation of PBMC using a density gradient, cryopreservation of PBMC, and thawing of cells as well as preparation for functional assays. For each protocol, we include important considerations, such as timing, storage temperatures, and freezing rate. In addition, we provide alternatives so that researchers can make informed decisions in determining the optimal protocol for their application.