Background & Aim
Leukapheresis products are commonly used as source material for cell and gene therapy
products. To avoid risks associated with shipping/storing fresh products and to optimize
manufacturing logistics, this starting material may be cryopreserved soon after collection.
Cryopreparation, the process of preparing cells for cryopreservation, typically involves
several manual steps including concentration/plasma depletion, optional washing/plasma
removal, and formulation with cryoprotectant. In this study, an automated system was
compared to manual processing for cryoprep of leukapheresis material using either
autologous plasma (AP) or synthetic solution (SYN), then frozen in either vials or
bags.
Methods, Results & Conclusion
Peripheral blood mononuclear cells (n=4) were collected from healthy donors, shipped
at 4°C to the processing facility, and processed within 24 hours. Products were split,
then processed manually via centrifugation in a test tube (MAN) or with a fully-automated
prototype small-volume processing system (AUTO, Fresenius Kabi). Cells were concentrated
and washed to <2e8 TNC/mL with either AP or SYN solution (2-8CELLsius, Protide Pharmaceuticals).
Suspensions were then diluted 1:1 with a fully-defined synthetic cryomedia (2-8CELLsius + 10%
DMSO, Protide Pharmaceuticals) to achieve a final concentration of 5% DMSO and <1e8
TNC/mL. The formulated cells were aliquoted into bags (20mL fill, AUTO samples) or
vials (1mL fill, all samples) and frozen at ∼-1°C/min in a controlled rate freezer.
Once frozen <-110°C, cells were immediately thawed in a 37°C bath. Samples were evaluated
for pre-freeze and post-thaw cell enumeration and viability (CD45+/7AAD−).
Hematology analyzer results showed no significant differences in TNC recovery between
cryopreparation method used (101.2% [±8.5%] MAN vs 102.2% [±4.4%] AUTO), wash solution
(101.2% [±5.6%] AP vs 102.4% [±6.4%] SYN), nor freezing format (104.2% [±3.3%] BAG
vs 100.6% [±6.6%] VIAL). Similarly, post-thaw viability showed no significant difference
between variables tested (84.4% [±7.1%] MAN vs 82.9% [±5.3%] AUTO vs 85.6% [±3.8%]
AP vs 81.1% [±6.8%] SYN vs 80.2% [±5.3%] BAG vs 84.9% [±5.6%] VIAL).
Leukapheresis cryoprep using a fully-automated system and synthetic wash solution
results in comparable post-thaw cell recovery and viability to manual processing with
autologous plasma.
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Copyright
© 2020 Published by Elsevier Inc.
