Page 110 - Eshre 2018

SLOW FREEZING PROTOCOLS – USE OF CRYOPROTECTIVE AGENTS
 Denmark: 1.5 mol/l ethylene glycol + 0.1 mol/l sucrose + 10 mg/ml HSA in PBS, tissue equilibrated for 30 minutes.
 Belgium: 1.5 mmol/L DMSO + 4 mg/mL of human serum albumin in Leibovitz medium, tissue equilibrated for 30 minutes
 Israel: 1.5 M DMSO + 15% synthetic serum substitute supplement + 0.1 M sucrose, tissue equilibrated for 30 minutes
 Germany: 10% CryoSure-DMSO + 10% serum substitute supplement in Leibovitz’s L-15 GlutaMAX medium, tissue equilibrated for 30 minutes
 Australia: 1.5 mol/l propanediol with 0.1 mol/l sucrose in base medium at room temperature for 90 min
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VITRIFICATION OF OVARIAN TISSUE (PROTOCOL: KAGAWA N, et al., 2009; SILBER S, et al., 2010)
Cortical tissue prepared 10mm x10mm Thickness: 1-1.5mm
Media 1: 7.5% EG + 7.5% DMSO+ 20% SSM for 25 min
Tissue placed on metal strip and plunged directly into liquid N2
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Media 2:
20% EG + 20% DMSO + 0.5M Sucrose for 15 min
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