Department of Transfusion Medicine and Hemostaseology, University Hospital Erlangen, Erlangen, Germany.
Transfusion. 2020 Jul;60(7):1500-1507. doi: 10.1111/trf.15756. Epub 2020 Mar 14.
Traditionally, white blood cells (WBCs) are collected from buffy coats or freshly drawn blood. However, the increasing demand for peripheral blood mononuclear cells (PBMCs) in the research phases of immunological therapy development makes it necessary to identify alternative sources of these cells.
Leukapheresis products are cost intensive and not offered by all blood banks. Therefore, thrombocyte apheresis cassettes (TACs), plateletpheresis waste products, were investigated as a possible low-cost and easily accessible blood source for research laboratories. The recovery rate, phenotype, and functionality of WBC subsets from TAC are unknown and were investigated in comparison to frequently used blood resources via flow cytometry.
On average, TACs provide 30.3 × 10 /mL PBMCs, situating themselves between peripheral whole blood (WB; 5.35 × 10 /mL) and leukoreduction system chamber (LRSC; 163.9 × 10 /mL) yields. Frequencies of CD14, CD3, CD4, CD8, CD56, CD19, and CD11c positive cells in TACs correlate with normal proportions of WBC populations. Stimulation of TAC-derived PBMCs by lipopolysaccharide (LPS) and resiquimod (R848) showed no significant differences in expression levels of human leukocyte antigen (HLA)-DR, DQ, DP, and CD86 or cytokine secretion compared to other blood source derived PBMC. Following stimulation with LPS or R848, comparable levels of tumor necrosis factor-α, interleukin-10, and interleukin-1β could be measured between TAC, LRSC, and WB. Additionally, TAC-derived T cells retained their proliferation capability and were able to produce interferon-γ following T-cell receptor stimulation.
TACs provide a cost-effective source of viable and functional human blood cells that can readily be used for clinical and laboratory investigations after plateletpheresis preparation.
传统上，白细胞（WBC）是从白细胞层或新抽取的血液中收集的。然而，外周血单个核细胞（PBMC）在免疫治疗开发的研究阶段的需求不断增加，这使得有必要寻找这些细胞的替代来源。
白细胞分离产品成本高昂，并非所有血库都提供。因此，血小板分离试剂盒（TAC），血小板分离废物，被认为是研究实验室低成本且易于获得的血液来源。TAC 中 WBC 亚群的回收率、表型和功能尚不清楚，通过流式细胞术与常用的血液资源进行了比较。
平均而言，TAC 提供 30.3×10 /mL 的 PBMC，位于外周全血（WB；5.35×10 /mL）和白细胞减少系统室（LRSC；163.9×10 /mL）之间。TAC 中 CD14、CD3、CD4、CD8、CD56、CD19 和 CD11c 阳性细胞的频率与 WBC 群体的正常比例相关。用脂多糖（LPS）和雷西莫德（R848）刺激 TAC 衍生的 PBMC 后，与其他血液来源的 PBMC 相比，人类白细胞抗原（HLA）-DR、DQ、DP 和 CD86 的表达水平或细胞因子分泌没有显著差异。用 LPS 或 R848 刺激后，TAC、LRSC 和 WB 之间可测量到相似水平的肿瘤坏死因子-α、白细胞介素-10 和白细胞介素-1β。此外，TAC 衍生的 T 细胞在 T 细胞受体刺激后保持增殖能力，并能够产生干扰素-γ。
TAC 是一种具有成本效益的人类活细胞来源，可在血小板分离准备后用于临床和实验室研究。
