SBIR Phase II: Dynamic Nanopores Enabling Non-toxic Cryopreservation to Advance Centralized Manufacturing & Transport of Regenerative Medicines

This Small Business Innovation Research Phase II project will continue development of cutting-edge nanopore technology that will greatly enhance biological preservation of regenerative medicines such as stem cells, complex tissues, and organs. Such a technology has the potential to greatly improve the biobanking and transport infrastructure of healthcare for on-demand cells and tissues, improved mass trauma care and advanced personalized medical procedures. Biopreservation is required in regenerative medicine at nearly all levels in the acquisition of source material, isolation, storage and shipment of a final product to patient. Yet critically, the field lacks the ability to safely and efficiently preserve these tissues and medicines while maintaining high cell viability and function after cryopreservation, severely limiting product shelf life. Nowhere is the absence of a biobanking technology more palpable than organ transplantation, where the time window between donor and recipient (4-7 hours) is not enough to properly match donations, screen for pathogens, or transport long distances. Enabling the United States to safely bank organs at subzero temperature will significantly enhance national healthcare. The US faces strong commercial and competitiveness reasons to invest in all facets of regenerative medicine, including organ therapies. Cryopreservation solutions would indirectly enable significant savings to the healthcare system, the patient, and healthcare insurance companies with the cost savings from regenerative medicine treatments estimated to be nearly $250 billion per year in the U.S. The cytotoxicity of current biopreservation techniques is largely associated with inefficient cryoprotective agent and water delivery across the cell membrane during cooling leading to irreparable cell damage from ice formation. Phase I established a fundamentally different approach to cryoprotective agent (CPA) optimization by developing first-in-the-field bioinspired nanopores as transmembrane mega highways to facilitate safe and efficient intracellular delivery and removal of cryoprotective agents during cryopreservation and dramatically increase post-thaw cell yield, viability, and function. The pore-formers assist CPA loading at reduced temperatures to make up the functional loss of biological protein channels at cold temperatures, reduce the amount of CPA required to remarkably low concentrations, and expand the selectable range of CPAs, all while dynamically dissociating at physiological temperature to prevent open-pore toxicity. Further R&D effort will diversify molecular design, optimize lead molecules, scale-up production, and perform cryopreservation studies on regenerative medicines of major therapeutic significance such as stem cells and complex tissues. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.