Kofanova Olga A, Davis Kristine, Glazer Barbara, De Souza Yvonne, Kessler Joseph, Betsou Fotini
1 Integrated BioBank Of Luxembourg (IBBL) , 6 rue Nicolas Ernest Barblé, L-1210, Luxembourg .
Biopreserv Biobank. 2014 Jun;12(3):206-16. doi: 10.1089/bio.2013.0090.
The impact of shipping temperatures and preservation media used during transport of either peripheral blood mononuclear cells (PBMCs) or Jurkat cells was assessed, in view of implementing of a proficiency testing scheme on mononuclear cell viability. Samples were analyzed before and after shipment at different temperatures (ambient temperature, dry ice, and liquid nitrogen) and in different preservation media (serum with cryoprotectant, commercial cryopreservation solution, and room temperature transport medium). Sample quality was assessed by viability assays (Trypan Blue dye exclusion, flow cytometry, Cell Analysis System cell counting (CASY)), and by ELISpot functional assay. The liquid nitrogen storage and shipment were found to be the most stable conditions to preserve cell viability and functionality. However, we show that alternative high quality shipment conditions for viable cells are dry ice shipment and commercial cryopreservation solution. These were also cost-efficient shipment conditions, satisfying the requirements of a proficiency testing scheme for viable mononuclear cells. Room temperature transport medium dramatically and adversely affected the integrity of mononuclear cells.
鉴于要实施一项关于单核细胞活力的能力验证计划，我们评估了运输外周血单核细胞（PBMC）或Jurkat细胞期间的运输温度和使用的保存介质的影响。在不同温度（环境温度、干冰和液氮）以及不同保存介质（含冷冻保护剂的血清、商用冷冻保存溶液和室温运输介质）下运输前后对样本进行分析。通过活力测定（台盼蓝染色排除法、流式细胞术、细胞分析系统细胞计数（CASY））以及ELISpot功能测定来评估样本质量。发现液氮储存和运输是保存细胞活力和功能最稳定的条件。然而，我们表明，对于活细胞而言，替代的高质量运输条件是干冰运输和商用冷冻保存溶液。这些也是具有成本效益的运输条件，满足了活单核细胞能力验证计划的要求。室温运输介质对单核细胞的完整性有显著的负面影响。
