Parallelized Droplet Vitrification for Single-Run Vitrification of Hepatocytes from an Entire Rat Liver

Abstract

Drug discovery pipelines rely on the availability of isolated primary hepatocytes for investigating potential hepatotoxicity prior to clinical application. These hepatocytes are isolated from livers rejected for transplantation and subsequently cryopreserved for later usage. The gold standard cryopreservation technique, slow-freezing, is a labor-intensive process with significant poststorage viability loss. In this work, we introduce parallelized droplet vitrification, a technique for high-volumetric, rapid vitrification of suspended cells. We show the utility of this technique through the single-run vitrification of the whole rat liver hepatocyte yield, resulting in the vitrification of 250 million cells in 40 mL of a vitrification solution at 10 mL/min. Additionally, we showed that these implementations maintained improved postpreservation outcomes in primary rat hepatocytes.