Abstract
Background & Aim Cryopreservation is a necessary step in the preparation of hematopoietic stem cells (HSC) used to reconstitute hematopoiesis for autologous stem cell transplantation. Cells are collected by leukapheresis, cryopreserved and stored while patients undergo preparative regimen. It is critical to optimize cryopreservation techniques to ensure optimal recovery of viable progenitor cells, especially when target cells are limited. Current methods are adequate, but limitations include inconsistency in components used to formulate media between institutions, increased risk for technical error and contamination. This study sought to optimize DMSO concentration and improve efficiency and compliance by comparing three different cryoprotectant media. Methods, Results & Conclusion Methods Ten HSC products, collected after standard mobilization of multiple myeloma patients, were cryopreserved with PRIME-XV FreezIS (Irvine Scientific) and compared to products previously cryopreserved with two standard use cryoprotectants: Std10 and Std5 (Std10 and Std5 are cryoprotectant media formulated in-house to achieve a final DMSO concentration of 10% and 5% respectively). Cells were processed and cryopreserved using institutional standard procedures, with storage for up to 6 months. At time of infusion, HSC were analyzed for CD34+ recovery and viability. Adverse events (AE) and time to engraftment were also monitored. Results Average CD34+ recovery for HSC cryopreserved with Std10, Std5 and PRIME-XV FreezIS were 39%, 74.7%, and 73.6% respectively (p-value: ≤ 0.001 Std10 vs. FreezIS). Median time to neutrophil engraftment was comparable (11 days) for all 3 media, while platelet engraftment occurred at a median of 20, 19 and 17 days respectively (p-value: >0.05 Std10 vs. FreezIS). Infusion-related AEs decreased from 80% (Std10) to 10% (FreezIS) (p-value: ≤ 0.0001). Discussion PRIME-XV FreezIS is a GMP-grade, pre-constituted cryoprotectant that minimizes cell processor variability and handling. In addition, compared to cryopreservation media prepared in house containing both 5% and 10% DMSO, PRIME-XV FreezIS results in ample CD34+ recovery and similar HSC engraftment. Finally, as anticipated, infusion toxicity caused by DMSO and frequency of AEs decreased significantly in patients infused with PRIME-XV FreezIS and Std5 when compared to products cryopreserved with Std10.
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