Viable Mononuclear Cell Stability Study for Implementation in a Proficiency Testing Program: Impact of Shipment Conditions

Abstract

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.

Biospecimen Science of Blood for Peripheral Blood Mononuclear Cell (PBMC) Functional Applications

“…Participation of PBMC processing laboratories in external quality assurance programs (EQA) is the only way to benchmark the performance of the laboratory. The benefits of participating in such programs have been demonstrated [79], although implementation of such a program requires optimization of the shipment conditions [80]. In conclusion, analysis of PBMC functional attributes requires standardization of the anticoagulant and blood collection tube type used, proper filling of the blood tubes, minimizing delay between blood collection and processing, and validation of the PBMC isolation protocol (centrifuge brand and settings, separation medium, buffer composition), the cryopreservation protocol (cryopreservation medium, device), the thawing protocol, and the post-thaw recovery.…”

A Biospecimen Proficiency Testing Program for Biobank Accreditation: Four Years of Experience

“…Cell viability is an important quality attribute of cryopreserved cells for many downstream applications such as functional tests, establishment of cell lines, or immunophenotyping. 22 The most common methods for cell viability assessment are trypan blue exclusion and flow cytometry. Both methods are based on plasma membrane integrity and use dyes that penetrate into cells with compromised membrane integrity (trypan blue dye) or intercalate DNA (propidium iodide).…”

Easy and cost-effective stable positioning of suspension cells during live-cell imaging

“…In order to investigate putative cytotoxic effects of the applied agarose pad on the cell suspension, a flow cytometry based viability testing [11] and a measurement of lactate dehydrogenase (LDH) release as a marker for membrane damage [12,13] were performed. Incubation of Jurkat cell for 2 h with an agarose pad showed a viability of over 87% while cells without an agarose pad had a viability of 95% (Fig.…”