As some plant species do not withstand the harsh conditions of cryopreservation and PVS treatment, a methodology was developed in which the explant follows an encapsulation process by ionic gelation. In this technique, an explant embedded in a drop of sodium alginate (2–3% v/w), the most studied encapsulating polymer, is placed with a pipet in a cationic solution, such as 100 mM calcium II chloride (CaCl2), for approximately 30 min to form protective capsules for the explants. A carrier solution (glycerol and/or sucrose) is also usually added to sodium alginate. These encapsulated explants are dehydrated—osmotically in a series of sucrose solutions (with increasing concentration; up to even 1.0 M), and physically with silica gel or in a laminar flow chamber (to a final level of 10–30% of the initial moisture content)—before immersion in LN [83,103].

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