The effect of cryopreservation on hMSC function

At Rooster Bio, Brooks Life Science Systems & BioLife Solutions

Cellular therapies have proven clinically effective and have the potential to revolutionize medicine. Cryopreservation in GMP-grade cryomedia and storage and transportation in vapor phase liquid nitrogen (LN2) are the preferred commercialization methods for cellular therapies. Routine sample access of cell banks stored in LN2 may expose adjacent doses to temperature excursions that could negatively impact the stability of frozen cells.

The purpose of this study was to assess the effect of two commonly employed cryopreservation medias and repetitive transient warming events (TWE) on clinically relevant mesenchymal stem/stromal cells (MSCs) and determine how the incorporation of evidence based Best Biopreservation Practices can improve cellular outcome.

Cryomedia composition impacts post-thaw hMSC functionality

Bar plots showing that cryomedia composition impacts post-thaw hMSC functionality

Because no TWE effect was observed in hMSCs after extended post-thaw culture, 0 and 20 TWE conditions were selected for assessment of short-term functionality. Cryopreserved hMSC exhibited similar recovery immediately post-thaw in all tested conditions as determined by membrane integrity using a NucleoCounter® NC-3000™. See figure: (A) viability; (B) recovery; and (C) viable recovery.

After thawing, hMSCs were transferred to appropriate culture vessels to facilitate cell adhesion and expansion. hMSCs were microscopically imaged prior to media change at two hours post-thaw. Figures represent mean + S.E.M (n = 6).

See full dataset in the PDF poster.


The data suggests that optimized cryopreservation can improve post-thaw hMSC function and reduce the need for extended post-thaw ‘rescue’ culture and may therefore improve the clinical efficacy of hMSC-based therapies.


Brian J. Hawkins, Alireza Abazari, Lye Theng Lock, Taby Ahsan, Jon A. Rowley, John Fink and Aby J. Mathew.