11. ガラス化法(vitrification)による植物培養細胞・組織の超低温保存(セミナー「低温環境への植物の適応-基礎と応用」)

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The development of a simple and reliable method for cryopreservation would allow much more widespread use of cryopreserved cultured plant cells and meristems. Potentially valuable cryogenic procedures would be vitrification and encapsulation/dehydration techniques. These new procedures increased the number of species to be cryopreserved over the last few years. In the vitrification method, cells and meristems are sufficiently dehydrated by a highly concentrated vitrification solution (PVS2) at 0 or 25℃. Thus, the procedure eliminates the need for controlled slow freezing and permits cells and meristems to be cryopreserved by direct transfer into liquid nitrogen. Successfully vitrified and warmed meristems directly developed shoots without inter-mediary callus formation. The encapsulation/dehydration technique is easy to handle and alleviates dehydration process. However, the problems are the lower rate of survival, the later recovery growth and the longer dehydration process compared with the vitrification procedure. To solve these problems, a novel cryogenic procedure, encapsulation/vitrification method, was presented. In this method, meristems were trapped into alginate-coated beads containing a mixture of 0. 4M sucrose and 2M glycerol. These encapsulated meristems (30-40) were dehydrated in a 100ml beaker containing 50 ml PVS2 for about 100 min at 0℃. This procedure is easy to handle and produces high levels of survival. Thus, the method promises for cryopreserving meristems, hairy roots, meristems and somatic embryos.
低温生物工学会の論文
1996-07-28

11. ガラス化法(vitrification)による植物培養細胞・組織の超低温保存(セミナー「低温環境への植物の適応-基礎と応用」)

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