Sakai A. auteur aut IRD Engelmann Florent auteur aut IRD text journalArticle eng text/pdf born digital access This paper discusses the importance of the successive steps of the vitrification technique and reviews the current development and use of vitrification and of the two derived protocols, encapsulation-vitrification and droplet-vitrification. Vitrification refers to the physical process by which a highly concentrated cryoprotective solution supercools to very low temperatures and finally solidifies into a metastable glass, without undergoing crystallization at a practical cooling rate. Samples are thus cryopreserved without detrimental intracellular ice formation. In a standard vitrification protocol, excised explants are precultured on medium enriched with sucrose, treated ('loaded') with a loading solution composed of 2 M glycerol + 0.4 M sucrose, dehydrated with a highly concentrated vitrification solution [e.g. the PVS2 vitrification solution, which contains 30% (w/v) glycerol, 15% (w/v) ethylene, glycol and 15% (w/v) DMSO and 0.4 M sucrose], frozen and rewarmed rapidly, unloaded with basal culture medium supplemented with 1.2 M sucrose, and then transferred to standard culture conditions. In the encapsulation-vitrification technique, the explants are encapsulated in alginate beads, loaded and dehydrated with a vitrification solution before rapid immersion in liquid nitrogen. In the droplet-freezing technique, excised explants are loaded, treated with the vitrification solution and frozen in individual microdroplets of vitrification solution placed on aluminium foils, which are immersed rapidly in liquid nitrogen. These three techniques have been applied to different tissues of over 100 plant species from temperate and tropical origins and the number of cases where they are being tested on a large scale or applied routinely is increasing. specialized plant germplasm long term conservation cryopreservation vitrification encapsulation vitrification droplet vitrification 076 28 3 151-172 2007 0143-2044 https://www.documentation.ird.fr/hor/fdi:010040731 0143-2044 [F B010040731] https://www.documentation.ird.fr/hor/fdi:010040731 https://www.documentation.ird.fr/intranet/publi/2007/10/010040731.pdf Accès réservé (Intranet de l'IRD) IRD - Base Horizon / Pleins textes 2007-11-06 2017-08-23 fdi:010040731 fre