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

Jinhao Hu; Xinqi Zhao; Guoying Bai

doi:10.1016/j.xpro.2023.102160

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

STAR Protoc. 2023 Mar 14;4(2):102160. doi: 10.1016/j.xpro.2023.102160. Online ahead of print.

Authors

Jinhao Hu¹, Xinqi Zhao¹, Guoying Bai²

Affiliations

¹ School of Materials Science and Engineering, Hebei University of Technology, Tianjin 300401, China.
² School of Materials Science and Engineering, Hebei University of Technology, Tianjin 300401, China; Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China. Electronic address: baiguoying@iccas.ac.cn.

Abstract

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.

Keywords: Cell Biology; Chemistry; Microscopy.