**2.5 Assessment of cell viability**

Cell viability was determined using the non-invasive metabolic indicator alamarBlue™ (Trek Diagnostics). A volume of 20 µl was added to cells in 200 µl of DMEM (10%FCS) and the plate was incubated at 37oC for 3 hours. Plates were read using a fluorescent microplate reader (Molecular Dynamics) at an excitation wavelength of 544 nm and an emission wavelength of 590 nm. Viability was measured before and after sugar loading, immediately after thawing and at several later time points post-thaw.

#### **2.6 Statistical methods**

All experiments were repeated at least four times with four replicates in each experiment. Statistical differences were assessed by two way analysis of variance. P-values 0.05 were regarded as significant.
