Abstract
Background aims
The use of effective methods for the cryopreservation of hematopoietic stem cells
(HSCs) is vital to retain the maximum engraftment activity of cord blood units (CBUs).
Current protocols entail the use of dimethyl sulfoxide (DMSO) as intracellular cryoprotective
agent (CPA) and dextran and plasma proteins as extracellular CPAs, but DMSO is known
to be cytotoxic, and its infusion in patients is associated with mild to moderate
side effects. However, new, commercially available, DMSO-free cryopreservation solutions
have been developed, but their capacity to protect HSCs remains poorly investigated.
Methods
Herein the authors compared the capacity of four DMSO-free freezing media to cryopreserve
cord blood (CB) HSCs: CryoProtectPureSTEM (CPP-STEM), CryoScarless (CSL), CryoNovo
P24 (CN) and Pentaisomaltose (PIM). Clinical-grade DMSO/dextran solution was used
as control.
Results
Of the four cryopreservation solutions tested, the best post-thaw cell viability,
recovery of viable CD45+ and CD34+ cells and potency were achieved with CPP-STEM,
which was equal or superior to that seen with the control DMSO. CSL provided the second
best post-thaw results followed by PIM, whereas CN was associated with modest viability
and potency. Further work with CPP-STEM revealed that CB CD34-enriched HSCs and progenitors
cryopreserved with CPP-STEM maintained high viability and growth expansion activity.
In line with this, a pilot transplantation assay confirmed that CPP-STEM-protected
CB grafts supported normal short- and long-term engraftment kinetics.
Conclusions
The authors’ results suggest that new, valuable alternatives to DMSO are now available
for the cryopreservation of HSCs and grafts, including CBUs.
Key Words
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Article info
Publication history
Published online: October 12, 2021
Accepted:
September 5,
2021
Received:
July 7,
2021
Identification
Copyright
© 2021 International Society for Cell & Gene Therapy. Published by Elsevier Inc. All rights reserved.
