Cryopreservation of olive embryogenic cultures.
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 degree C per min). 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.
Sánchez-Romero
C
C
IFAPA Centro de Churriana, Churriana-Malaga, SPAIN. c.sanchez@uma.es
Swennen
R
R
Panis
B
B
D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000009:adverse effects / Q000494:pharmacology; D004121:Dimethyl Sulfoxide / Q000009:adverse effects / Q000494:pharmacology; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology; D019855:Ethylene Glycol / Q000009:adverse effects / Q000494:pharmacology; D005990:Glycerol / Q000009:adverse effects / Q000494:pharmacology; D031658:Olea / Q000187:drug effects / Q000196:embryology* / Q000502:physiology*; D013395:Sucrose / Q000009:adverse effects / Q000494:pharmacology
D003451:Cryoprotective Agents; D013395:Sucrose; D019855:Ethylene Glycol; D005990:Glycerol; D004121:Dimethyl Sulfoxide