McQuillan Adrian, Dotti Silvia, Cristiano Antonio, Ferrari Maura, Razzuoli Elisabetta, Amadori Massimo
AM Robotic Systems Limited, 10 Cranleigh Close, Warrington WA4 6SD, UK.
Biologicals. 2012 Jan;40(1):31-5. doi: 10.1016/j.biologicals.2011.09.009. Epub 2011 Oct 19.
Investigations into immune responses are often based upon recovery of peripheral blood mononuclear cells (PBMC). To this purpose, the recovery of PBMC by gradient centrifugation is labour-intensive and requires a reasonable level of skill by the laboratory technician. Thus, we set out to determine whether laboratory automation equipment could be used for the recovery of PBMC from blood samples of horses, pigs and cattle, based on the Ficoll-Paque gradient centrifugation technique. Mixing of blood samples with PBS, layering of diluted blood onto Ficoll-Paque gradients, recovery of separated PBMC in RPMI 1640 medium were performed using an automated robotic system, the SBF200 (AM Robotic Systems, Warrington, UK) under laminar air flow conditions. Tubes were tagged with bar codes and manually placed after gradient centrifugation into a tube reader to measure the volume and position of the PBMC layer. The results of the automated procedure compared very well to those of the manual one in terms of percent cell recovery, sterility and cell viability. Also, a high throughput of samples could be implemented: with the integration of cell counting it should be possible for 96 blood samples to be processed, including the production of aliquots, by one person in a day.
对免疫反应的研究通常基于外周血单核细胞（PBMC）的回收。为此，通过梯度离心回收PBMC既耗费人力，又要求实验室技术人员具备一定水平的技能。因此，我们着手基于Ficoll-Paque梯度离心技术，确定实验室自动化设备是否可用于从马、猪和牛的血液样本中回收PBMC。在层流空气流动条件下，使用自动化机器人系统SBF200（AM Robotic Systems，英国沃灵顿）将血液样本与PBS混合，将稀释后的血液铺在Ficoll-Paque梯度上，在RPMI 1640培养基中回收分离的PBMC。用条形码标记试管，并在梯度离心后手动放入试管读取器中，以测量PBMC层的体积和位置。在细胞回收率、无菌性和细胞活力方面，自动化程序的结果与手动程序的结果非常吻合。此外，还可以实现高通量样本处理：通过整合细胞计数，一个人一天应该能够处理96个血液样本，包括制备等分试样。
