Human neuronal cell viability demonstrated in culture after cryopreservation

Cryopreservation of transfected primary dorsal root ganglia neurons

“…The concentration of cryoprotectants can also affect survival rates of cryopreserved cells. The amount of cryoprotectant (DMSO) used has been reported between 5-10% DMSO (Silani et al, 1988;Ruwe and Trumble, 1990;Petite and Calvet, 1995;Negishi et al, 2002;Ma et al, 2006). In these cases, maximal recovery occurred with the use of 8-10% DMSO, which is consistent with the present conditions (Silani et al, 1988;Negishi et al, 2002).…”

“…In our study, dissociated neurons were immediately placed in Styrofoam at −80 • C up to a week before being transferred to liquid nitrogen. Cryopreservation protocols, however, note the importance of using a freezing container or stepwise freezing conditions to provide a freezing rate of 1 • C/min, allowing the cryoprotectant to reach equilibrium within the cell (Silani et al, 1988;Ruwe and Trumble, 1990;Petite and Calvet, 1995;Negishi et al, 2002;Ma et al, 2006). Additionally, allowing for a recovery period prior to cryopreservation is another parameter which can potentially be optimized to increase viability post-cryopreservation.…”

The development and characterisation of complex ovine neuron cultures from fresh and frozen foetal neurons

“…Viability of the thawed cells, 23-48%, was within 12-60% range reported for freezing rat (Mattson andKater, 1988, Swett et al, 1994) and human cells (Silani et al, 1988) by various methods, and much better than obtained using more sophisticated programmed time-dependent temperature reductions in cryopreservatives. Neuron cultures from frozen stocks and freshly cultured cells contained similar proportions of the various neuron types.…”

Cryopreservation of GABAergic Neuronal Precursors for Cell-Based Therapy

“…Dimethyl sulfoxide (Me 2 SO), the most extendedly used intra-cellular cryoprotectant, has been reported to maintain multipotency and render excellent viability of murine neural progenitors when applied at 7–10% [23]. Based on the Me 2 SO usage background and the previous literature about nervous tissue and neural progenitor cryopreservation [24–29], we considered this crioprotectant as likely suitable to preserve MGE-derived GABAergic neuronal precursors. Regarding cooling rates, there are two general methods.…”