Cryopreservation of undifferentiated and differentiated human neuronal cells

The effective use of human-derived cells that are difficult to freeze, such as parenchymal cells and differentiated cells from stem cells, is crucial. A stable supply of damage-sensitive cells, such as differentiated neuronal cells, neurons, and glial cells can contribute considerably to cell therapy. We developed a serum-free freezing solution that is effective for the cryopreservation of differentiated neuronal cells. The quality of the differentiated and undifferentiated SK-N-SH cells was determined based on cell viability, live-cell recovery rate, and morphology of cultured cells, to assess the efficacy of the freezing solutions. The viability and recovery rate of the differentiated SK-N-SH neuronal cells were reduced by approximately 1.5-folds compared to that of the undifferentiated SK-N-SH cells. The viability and recovery rate of the differentiated SK-N-SH cells were remarkably different between the freezing solutions containing 10% DMSO and that containing 10% glycerol. Cryoprotectants such as fetal bovine serum (FBS), antifreeze proteins (sericin), and sugars (maltose), are essential for protecting against freeze damage in differentiated neuronal cells and parenchymal cells. Serum-free alternatives (sericin and maltose) could increase safety during cell transplantation and regenerative medicine. Considering these, we propose an effective freezing solution for the cryopreservation of neuronal cells. (C) 2022, The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.

Automated summary

The effective cryopreservation of difficult-to-freeze cells, such as differentiated neuronal cells, is crucial for cell therapy. We developed a serum-free freezing solution that showed promising results in preserving differentiated neuronal cells. Evaluation of the freezing solutions was based on cell viability, recovery rate, and morphology. Cryoprotectants like fetal bovine serum, antifreeze proteins, and sugars were found to be essential for protecting against freeze damage.