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Cellular Encapsulation in 3D Hydrogels for Tissue Engineering
Published on: October 26, 2009
Cryopreservation of apple (Malus×domestica Borkh.) shoot tips following encapsulation-dehydration or
H Paul1, G Daigny1, B S Sangwan-Norreel1
1Faculté des Sciences, Laboratoire Androgenèse et Biotechnologie, 33 rue Saint Leu, 80039 Amiens Cedex 01, France Fax: +33-0322-827612 e-mail Brigitte.Sangwan@sc.u-picardie.fr, , , , , , FR.
Abstract:
Axillary shoot tips of apple cv. Golden Delicious isolated from shoot cultures were successfully cryopreserved using the encapsulation-dehydration technique. After encapsulation in alginate gel, embedded shoot tips were dehydrated by exposure to a sterile air flow before being frozen in liquid nitrogen and subsequent slow thawing. A preculture on modified MS medium containing 0.75 M sucrose followed by 6 h of dehydration (21% residual water) led to the highest shoot regrowth of frozen, coated shoot tips (83.7%). Among the sugars tested, sucrose and sorbitol presented the best cryoprotective effect. Four other scion apple varieties and rootstocks were also successfully cryopreserved. Axillary shoot tips of five apple (Malus×domestica Borkh.) scion and rootstock cultivars were cryopreserved using the encapsulation-vitrification technique. Using a one-step freezing method, we successfully cryopreserved axillary shoot tips without the requirement of a cold hardening pretreatment of the shoot cultures. Cryopreserved shoot tips treated with aqueous cryoprotective mixture IV containing 180% (w/v) sucrose and 120% (v/v) ethylene glycol showed the highest shoot regrowth rates, which varied from 64% to 77%, depending on the cultivar.
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