Aby J. Mathew

Cryopreservation of umbilical cord blood with a novel freezing solution that mimics intracellular ionic composition.

BACKGROUND: Cryopreservation protocols have remained relatively unchanged since the first umbilical cord blood banking program was established. This study evaluated the preservation efficacy of a novel intracellular‐like cryopreservation solution (CryoStor, BioLife Solutions, Inc.), the rate of addition of two cryopreservation solutions to cord blood units (CBUs), and reduced final dimethyl sulfoxide (DMSO) concentration of 5%.

Clinical methods of cryopreservation for donor lymphocyte infusions vary in their ability to preserve functional T-cell subpopulations

Cryopreserved donor lymphocyte infusion (DLI) products are manufactured and administered to treat relapse after allogeneic hematopoietic stem cell transplantation. Reported clinical responses to DLIs vary broadly, even within the same group of patients. While there is an implicit recognition of the fact that different manufacturing protocols may have specific effects on different cell types, cryopreservation protocols are frequently derived from our experience in the cryopreservation of stem cell products and do not account for the heterogeneous functional nature of DLI T‐cell populations. Here, we report the results of a prospective, multicenter trial on the effect of four different cryopreservation solutions that were used to freeze DLIs compared to control DLIs that were refrigerated overnight.

The effect of cryomedia selection and transient warming events on post-cryopreservation human MSC function

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 TWE on clinically-relevant Mesenchymal Stem/Stromal Cells (MSCs), and determine how the incorporation of evidence-based Best Biopreservation Practices can improve cellular outcome. Xeno-free human MSCs (RoosterBio, Frederick, MD) were cryopreserved either in a traditional home-brew MSC cryomedia (HB; 5% Human Serum Albumin/10% DMSO/Plasma-Lyte A) or in GMP-manufactured CryoStor CS5Ò cryomedia (CS5; BioLife Solutions, Bothell, WA). To mimic clinical conditions, cryopreserved samples were shipped in an evoÒ DV-4 LN2 smart shipper (Savsu Technologies, Albuquerque, NM) to Brooks Life Sciences (Chelmsford, MA). Upon receipt, samples were divided into separate cryoboxes for each experimental condition and stored in a LN2 vapor freezer (BioStore III Cryo automated freezer). Over two weeks, the cryoboxes were removed from the freezer to warm the samples to approximately -110°C for 0, 5, 10, 15, and 20 TWEs. Samples were then sent to RoosterBio and BioLife for determination of viability, expansion, IDO secretion, and metabolic activity both immediately post-thaw and following post-thaw culture. Immediate post-thaw viability was similar between samples frozen in HB and CS5 with no overall trend observed between viability and TWE number. All cells experienced rapid post-thaw cell expansion in culture. Despite these apparent similarities, cellular metabolic activity (alamarBlue) was elevated in cells frozen in CS5 versus HB both immediately post-thaw and after 24 hr of culture, and a decline in MSC immunomodulatory activity was apparent at 20 TWE. These results suggest that both the cryomedia employed and the number of temperature excursions during storage affect functionality and potency of cryopreserved cellular therapies. As such, our findings emphasize the importance of optimized cryopreservation protocols and storage conditions to maximize the effectiveness of cellular therapies in patients. Brian J. Hawkins1,2, Alireza Abazari1, Lye Theng Lock3, Taby Ahsan3, Jon A. Rowley3, John Fink4, Aby J. Mathew1