Protocol for cryopreservation of red blood cells that controls ice crystal formation by fulvic acid

Abstract

Summary: The development of biocompatible ice-controlling materials for non-vitreous cryopreservation of cells is of great importance to the field of biomedicine. Here, we present a protocol to use fulvic acid (FA) for efficient non-vitreous cryopreservation of red blood cells (RBCs) that both promotes the melting of ice crystals and retards their growth/recrystallization. We describe steps for FA fractionation and performing tests for ice recrystallization and ice freezing/thawing. We then detail the freezing/thawing of RBCs, recovering RBCs, and testing their viability.For complete details on the use and execution of this protocol, please refer to Bai et al. (2022).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Keywords

• Cell Biology
• Microscopy
• Chemistry