Direct comparison of Cryotop® vitrification and Bicell® freezing on recovery of functional rat pancreatic islets.

Two protocols, Bicell® freeze-thawing and Cryotop® vitrification-warming, were compared for suitability in cryopreserving rat pancreatic islets (101-150 μm in mean diameter). Immediate survival rates of post-thaw and post-warm islets (50 and 57%, respectively), assessed by FDA/PI double staining, were lower than that of fresh control islets (90%). Most of the PI-positive dead cells were detected in peripheral area of post-warm islets, and were removed after subsequent 24 h culture (survival rate; 85% vs 59% in post-thaw islets). Quantitative PCR analysis showed that Bicell® freeze-thawing compromised expression of genes relating to β-cell function (Pdx1 and Glut2), but not to one of apoptotic pathways (Bax/Bcl2 ratio). Expression of these genes was maintained in islets before and after the Cryotop® vitrification-warming. Values of stimulus index (SI) for 20 mM/3 mM glucose-stimulated insulin secretion were 6.7, 1.9 and 3.9 in fresh control, post-thaw and post-warm islets, respectively. The SI values after 24 h culture were 4.1, 1.9 and 3.1, respectively. Larger islets (>150 μm in diameter) had comparable survival rates, but lower SI values after Cryotop® vitrification-warming when compared to smaller counterparts. These results suggest that rat pancreatic islets can be cryopreserved by Cryotop® vitrification-warming rather than Bicell® freeze-thawing, without considerable loss of in vitro β-cell function.