Knörck Arne, Marx Stefanie, Friedmann Kim S, Zöphel Sylvia, Lieblang Lisa, Hässig Carmen, Müller Isabelle, Pilch Jan, Sester Urban, Hoth Markus, Eichler Hermann, Sester Martina, Schwarz Eva C
Biophysics, Center for Integrative Physiology and Molecular Medicine (CIPMM), School of Medicine, Homburg, Germany.
Department of Transplant and Infection Immunology, Saarland University, Homburg, Germany.
Transfusion. 2018 Jun;58(6):1516-1526. doi: 10.1111/trf.14616. Epub 2018 May 6.
Research with primary human white blood cell (WBC) subpopulations requires high quantity, quality, and functionality of peripheral blood mononuclear cells (PBMCs) as a source to further characterize cellular subpopulations such as T and B lymphocytes, monocytes, or natural killer cells. Apart from buffy coats derived from whole blood, residual blood from preparative hemapheresis kits are used as a source for PBMCs, but knowledge on the yield and functionality of cells from different devices is limited.
We evaluated quantity and quality of PBMCs isolated from apheresis kits of two apheresis devices (AMICUS, Fenwal; and Trima Accel, Terumo BCT), the latter being our standard source for many years. PBMCs derived from Trima or AMICUS were tested for yield and subtype composition by flow cytometry. Functionality was assessed by cytokine induction of CD4 and CD8 T cells and by degranulation. Moreover, cytotoxic activity of natural killer cells was quantified by a real-time killing assay.
Mean numbers of isolated cells were 5.5 ± 2.4 × 10 for AMICUS, and 10.3 ± 6.4 × 10 for Trima Accel, respectively. The proportion of WBC subtypes corresponded to well-known numbers from whole blood, with minor differences between the two apheresis systems. Likewise, minor differences in cytokine induction were found in stimulated CD4 or CD8 T cells. Finally, PBMCs derived from the two systems showed comparable cytotoxic activity.
PBMC derived from residual blood of the AMICUS and Trima Accel apheresis devices serve as an economic and easily accessible source for functional PBMCs with comparable quantity and quality to PBMCs derived from whole blood.
对原代人白细胞(WBC)亚群进行研究需要大量、高质量且具有功能的外周血单核细胞(PBMC)作为来源,以进一步表征细胞亚群,如T和B淋巴细胞、单核细胞或自然杀伤细胞。除了来自全血的血沉棕黄层外,制备性血液成分单采试剂盒中的残余血液也被用作PBMC的来源,但关于不同设备中细胞产量和功能的知识有限。
我们评估了从两种血液成分单采设备(Fenwal公司的AMICUS和Terumo BCT公司的Trima Accel)的血液成分单采试剂盒中分离出的PBMC的数量和质量,后者多年来一直是我们的标准来源。通过流式细胞术检测从Trima或AMICUS获得的PBMC的产量和亚型组成。通过CD4和CD8 T细胞的细胞因子诱导以及脱颗粒来评估功能。此外,通过实时杀伤试验对自然杀伤细胞的细胞毒性活性进行定量。
AMICUS分离出的细胞平均数量为5.5±2.4×10,Trima Accel分离出的细胞平均数量为10.3±6.4×10。白细胞亚型的比例与全血中众所周知的数值相符,两种血液成分单采系统之间存在细微差异。同样,在受刺激的CD4或CD8 T细胞中发现细胞因子诱导存在细微差异。最后,来自这两种系统的PBMC表现出相当的细胞毒性活性。
来自AMICUS和Trima Accel血液成分单采设备残余血液的PBMC可作为一种经济且易于获取的功能性PBMC来源,其数量和质量与来自全血的PBMC相当。