Enhanced Cryopreservation of Therapeutically Relevant Cells

Published on 13 May 2026

Superior Performance of Recombinant Albumin and Its Role in Reducing DMSO Dependency

Reduced-DMSO cryopreservation workflows often compromise post-thaw viability, expansion, and functional cell attributes due to increased cellular stress during freeze/thaw. In a comparative study across T cells, mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSCs), InVitria® evaluated whether recombinant human serum albumin (rHSA) could improve cryopreservation performance relative to plasma-derived HSA (pHSA) and albumin-free controls across 2%, 5%, and 10% DMSO formulations. Post-thaw recovery, apoptosis, proliferation, viability, and pluripotency were assessed alongside analytical characterization of albumin aggregation and Cys34 redox state. Recombinant albumin supported maintained or improved post-thaw performance at reduced DMSO concentrations while demonstrating significantly lower aggregate burden and improved redox consistency compared to plasma-derived material.

Key Findings

The study established several findings with direct implications for cell therapy cryopreservation and manufacturing workflows:

• Recombinant albumin supported reduced-DMSO cryopreservation while maintaining or improving post-thaw recovery across T cells, MSCs, and iPSCs
• In T cells, 5% DMSO with rHSA matched the expansion performance of conventional 10% DMSO formulations
• In MSCs, rHSA reduced apoptosis and preserved post-thaw viability under low-DMSO conditions
• In iPSCs, rHSA improved recovery and expansion while preserving pluripotency markers at reduced DMSO concentrations
• Analytical characterization showed recombinant albumin contained substantially lower aggregate levels and more consistent Cys34 redox profiles than plasma-derived HSA

Why Cryopreservation Formulations Matter

Cryopreservation is a critical step in cell therapy manufacturing, enabling cell storage, transport, process flexibility, and final drug product preparation. However, conventional cryopreservation systems frequently rely on high concentrations of dimethyl sulfoxide (DMSO), which is associated with cellular toxicity, apoptosis, altered phenotype, and downstream patient safety concerns.

Albumin is commonly incorporated into cryopreservation formulations because it provides multiple protective functions during freeze/thaw stress. Albumin reduces ice crystal formation by interacting with water molecules, mitigates oxidative stress through antioxidant activity, and helps preserve membrane integrity and cellular viability during freezing and thawing.

Plasma-derived human serum albumin has historically been used for this purpose, but donor-derived material introduces structural heterogeneity, oxidation, aggregate burden, and lot-to-lot variability that may affect reproducibility in sensitive workflows. Recombinant albumin eliminates many of these liabilities while maintaining the functional benefits associated with albumin supplementation.

Study Design

T cells, MSCs, and iPSCs were expanded and cryopreserved using formulations containing 2%, 5%, or 10% DMSO combined with recombinant human serum albumin, plasma-derived HSA, or no albumin controls. Following thaw, cells were monitored using live-cell imaging systems to evaluate proliferation, viability, apoptosis, and functional recovery over multiple days in culture.

Additional analytical characterization compared recombinant and plasma-derived albumin using native capillary electrophoresis to quantify aggregate burden and reverse-phase HPLC to assess Cys34 redox state and oxidative status.

Results

Improved Post-Thaw T Cell Expansion at Reduced DMSO

Recombinant albumin improved post-thaw T cell expansion across multiple DMSO concentrations. Notably, formulations containing 5% DMSO with rHSA achieved expansion performance comparable to standard 10% DMSO conditions, supporting equivalent growth with reduced cryoprotectant exposure. See the full Application Note.

Reduced MSC Apoptosis and Maintained Viability

In MSC workflows, recombinant albumin reduced apoptosis and restored post-thaw viability under reduced-DMSO conditions. Albumin-free controls showed substantially lower viability and increased apoptosis, particularly at lower DMSO concentrations. See the full Application Note.

Enhanced iPSC Recovery and Preservation of Pluripotency

In iPSCs, recombinant albumin improved post-thaw recovery and expansion while preserving pluripotency markers under reduced-DMSO conditions. Formulations containing rHSA outperformed albumin-free controls and matched or exceeded plasma-derived HSA performance across multiple endpoints. See the full Application Note.

Structural Homogeneity Supports Functional Performance

Analytical characterization demonstrated that recombinant albumin contained substantially lower aggregate levels and more reduced Cys34 species compared to plasma-derived HSA. Recombinant albumin also demonstrated significantly lower lot-to-lot variability across tested batches, supporting more predictable cryopreservation performance. See the full Application Note.

Conclusion

Recombinant human serum albumin supports reduced-DMSO cryopreservation while maintaining or improving post-thaw viability, expansion, and functional cell attributes across multiple therapeutically relevant cell types. By combining lower aggregate burden, improved redox consistency, and animal-origin-free production, recombinant albumin provides a more controlled and reproducible alternative to plasma-derived HSA for advanced cell therapy manufacturing workflows.

Download the Poster

Download the full scientific poster presentation: Enhanced Cryopreservation of Therapeutically Relevant Cells: Superior Performance of Recombinant Albumin and Its Role in Reducing DMSO Dependency.