Novel Biocompatible, High-Throughput Cryopreservation Medium and Method for Enhanced Post-Thaw Migration and Cytotoxicity of NK Cells

SUMMARY Recent breakthroughs in CAR-T cell therapies have revolutionized immunotherapy, sparking growing interest in NK cell-based treatments due to their lower risk of graft-versus-host disease and ability to target tumors independently of MHC class I expression. However, current cryopreservation methods using toxic, cell- permeating agents significantly impair NK cell migration, cytotoxicity, and cytokine production, limiting their therapeutic efficacy and scalability for clinical applications. CryoCrate, an NIH Innovation Showcase Company, is developing OdinSol®, a novel polysaccharide-based cryopreservation medium specifically designed to enhance the post-thaw viability and functionality of NK cells without the use of permeating cryoprotectants. OdinSol® leverages nanoscale cubic ice formation and molecular assembly technologies to preserve NK cell migration, cytotoxicity, and cytokine production. This project aims to achieve three milestones: 1. Optimize OdinSol® for primary NK cells, ensuring high post-thaw viability, enhanced cytotoxicity, and validated tumor-killing efficiency in vitro. 2. Produce a GMP-compliant prototype batch with verified stability and quality control, ensuring the scalability of OdinSol® production. 3. Validate the in vivo safety and efficacy of GMP-grade OdinSol® in immunotherapy applications by using a standard animal model, and secure FDA clearance for clinical use. The success of this project will significantly improve the manufacturing and clinical application of NK cell therapies, making them more scalable, effective, and accessible for cancer treatment.