Leukoreduction system chambers are an efficient, valid, and economic source of functional monocyte-derived dendritic cells and lymphocytes
Leukoreduction system chambers are an efficient, valid, and economic source of functional monocyte-derived dendritic cells and lymphocytes
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DOI：
10.1016/j.imbio.2013.07.005
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发表时间：
2013-11-01
期刊：
影响因子：
2.8
通讯作者：
Knippertz, Ilka
中科院分区：
文献类型：
--
作者：
Pfeiffer, Isabell A.;Zinser, Elisabeth;Knippertz, Ilka
近年来，对人类单核细胞来源的树突状细胞（moDCs）以及人类原代B淋巴细胞和T淋巴细胞用于免疫学研究的需求有所增加。通常，这些单核细胞是通过外周血单个核细胞（PBMCs）的塑料粘附从健康供者的血液、白细胞分离产物或白皮肤中分离出来的，然后用粒细胞巨噬细胞集落刺激因子（GM-CSF）和白细胞介素(IL)-4刺激，而淋巴细胞通常通过磁性细胞分选从非粘附部分（NAF）中分离出来。然而，献血者的血液是一种有限的资源，并不是每个血库都提供白细胞分离产品或实验室使用的暗色外套。此外，白细胞分离是非常昂贵的，而且细胞的产生/分离是时间和成本密集的。为了克服这些障碍，我们评估了低成本的白细胞诱导系统腔（LRSCs）是否可以作为生成modc和分离淋巴细胞的PBMCs的替代和适当来源。LRSCs是在常规供体血小板提取过程中产生的，通常被丢弃。通过分析来自LRSCs的未成熟和成熟树突状细胞（dc）以及B淋巴细胞和T淋巴细胞的数量和表型，我们发现所有细胞的数量和质量都很高。对dc的进一步研究包括跨井迁移试验、异体混合淋巴细胞反应（MLR）、细胞因子分泌试验和细胞毒性T细胞诱导试验，结果显示这些细胞具有高迁移性和刺激能力。此外，dc和T细胞被mRNA有效电穿孔，并在共培养后显示出特征性的细胞因子产生，证明LRSCs是一种高效、有效和经济的来源，用于研究目的的modc和淋巴细胞的产生。(C) 2013爱思唯尔有限公司版权所有。
The demand for human monocyte-derived dendritic cells (moDCs), as well as for primary human B and T lymphocytes for immunological research purposes has been increased in recent years. Classically, these monocytes are isolated from blood, leukapheresis products or buffy coats of healthy donors by plastic adherence of peripheral blood mononuclear cells (PBMCs), followed by stimulation with granulocyte macrophage colony-stimulating factor (GM-CSF) and interleukin (IL)-4, while lymphocytes are usually isolated from the non-adherent fraction (NAF) by magnetic cell sorting. However, donor-blood is a limited resource and not every blood bank offers leukapheresis products or buffy coats for laboratory use. Additionally, a leukapheresis is very expensive and also the generation/isolation of cells is time- and cost-intensive. To overcome some of these obstacles, we evaluated if low-cost leukoreduction system chambers (LRSCs), which arise after routine donor plateletpheresis procedures, and are usually discarded, would be an alternative and appropriate source of PBMCs to generate moDCs and to isolate lymphocytes. By analyzing the number and phenotype of immature and mature dendritic cells (DCs), as well as of B and T lymphocytes derived from LRSCs, we found all cells to be of high quantity and quality. Further investigations on DCs comprising transwell migration assays, allogeneic mixed lymphocyte reactions (MLR), cytokine secretion assays, and cytotoxic T cell induction assays revealed high migratory, as well as stimulatory capacity of these cells. In addition, DCs and T cells were efficiently electroporated with mRNA and showed characteristic cytokine production after co-culture, demonstrating LRSCs as an efficient, valid, and economic source for generation of moDCs and lymphocytes for research purposes. (C) 2013 Elsevier GmbH. All rights reserved.