We established a simple cryopreservation protocol for Arabidopsis T87 cells using an encapsulation-dehydration method. T87 cells were encapsulated into alginate beads containing 2 M glycerol and 0.4 M sucrose. Alginate beads containing T87 cells were dehydrated with silica gel for 2 h (to c. 0.7 g H2O g DW-1) followed by immersed in LN. After rewarming at 35°C for 3 min and 1-d incubation under continuous illumination at 22°C, cryopreserved T87 cells exhibited considerable regrowth. Exponentially-grown 7-d-old T87 cells regrew more vigorously (86% of control) than 14-d-old cells after cryopreservation without preculture in medium containing 0.3 M sucrose. Genetic stability of cryopreserved T87 cells was demonstrated by gas chromatography time-of-flight mass spectrometry (GCT-OF-MS) and principal component analysis (PCA). Transformed T87 cells were cryopreserved using established protocols, and GUS expression was maintained within a 2-fold variance. These results indicate that cryopreservation of T87 cells is useful for comprehensive metabolomics research and for the large scale collection of transformed cultured cell lines for functional genomics research.
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Document Type: Research Article
Publication date: 01 September 2008
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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.