Abstract
-1 the viability on thawing increased from 39.9 ± 2.0% in nonnucleated sample to 65.9 ± 4.5% followed controlled ice nucleation and a consequent increase in hESC colony survival following 48 hr culture. Electron micrographs (CryoSEM and freeze-substitution) of hESC in the frozen state are presented to illustrate the beneficial effects of ice nucleation on the osmotic shrinkage of hESC during cooling and the subsequent avoidance of intracellular ice. Efficient nucleation is essential for high levels of hESC viability The EF600 was used to freeze hESC in a linear freeze profile, using Cryostor CS10, with an ice nucleation step at 9.0°C where indicated. Efficient nucleation of the vials resulted in an improvement of immediate viability of 26% compared to that of non-nucleated cells (from 39.9 ± 2.0% to 65.9 ± 4.5%) and a consequent increase in hESC colony survival following the 48 hr culture period (figure 1).
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