The aim of this work was to optimize a protocol for the cryopreservation of embryogenic cultures of olive (Olea europaea L.). Exposure time to loading solution and PVS2 significantly influenced the regrowth rate of both organized and non-organized tissues. Organized tissues were more sensitive to prolonged treatments with vitrification solutions compared to non-organized tissues. Three cryopreservation protocols were compared using non-organized tissues: the "classical" vitrification protocol, an ultra-fast freezing method using droplet vitrification on aluminium foil strips and a "classical" slow freezing method (1°C min−1). The best results were obtained using the droplet vitrification method after a 60 min dehydration period with PVS2. Under these conditions, all cryopreserved cultures showed renewed embryogenesis six weeks after thawing. A long-term (7-8 weeks) sucrose preculture had a significant effect on the initial response of the cultures, allowing particularly to protect cells against the toxic effects of the vitrification solution.
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Document Type: Research Article
Publication date: September 1, 2009
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CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation
The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.