A multistage-dialysis microdevice for extraction of cryoprotectants

Abstract

In this study, we present a multistage-dialysis microdevice (MDM) for extraction of cryoprotectants (CPAs) from a CPA-laden cell suspension. We confirmed the functions of the key designs of the MDM using a fluorescence solution, we assessed the performance of the MDM by using the MDM to unload glycerin from glycerin-loaded swine erythrocytes, and we investigated the effects of the cell suspension flow rate, glycerin concentration, cell density, and membrane pore size on the clearance efficiency of glycerin (C), the survival rate of cells (S), and the recovery rate of cells (R). Under the designed conditions, C, S, and R reached ~60%, ~90%, and ~70%, respectively. In addition, a high flow rate causes high S and R but a low C. For a low glycerin concentration, C, S, and R are all high. If a low cell density or a large pore membrane is used, C is high, whereas both S and R are low. This work provides insight into the development of microfluidic devices for the inline extraction of cryoprotectants from a small volume of cryopreserved cells prior to the use of the cells in lab-on-a-chip applications.