Anthony P, Lowe KC, Power JB, Davey MR
Department of Life Science, University of Nottingham, Nottingham, NG7 2RD, United Kingdom.
Cryobiology. 1997 Nov;35(3):201-8. doi: 10.1006/cryo.1997.2041.
The beneficial effects were assessed of supplementing culture medium with oxygenated perfluorocarbon, both alone and in combination with 0.01 (w/v) Pluronic F-68, on the postthaw viability, following cryopreservation, of suspension cultured cells of the Japonica rice, Oryza sativa cv. Taipei 309. The mean viability, as assessed by triphenyl tetrazolium chloride reduction, of cells at 4 days after thawing was increased 20% over control by oxygenated perfluorodecalin (P < 0.05) in 100-ml glass jars and similarly by 24% (P < 0.01) when recovered on a smaller scale in 24-well petri dishes. In a separate assessment, a 21% increase above control treatments (P < 0.05) in postthaw viability was also observed with oxygenated perfluorodecalin. A similar, 36% increase (P < 0.05) in postthaw viability occurred with cells exposed to 0.01% (w/v) Pluronic F-68 alone. A more pronounced, synergistic increase in viability, up to 57% over control (P < 0.05), occurred with cells recovered in the presence of both oxygenated perfluorodecalin and 0. 01% (w/v) Pluronic F-68. No significant difference was observed between Pluronic F-68 and oxygenated perfluorodecalin treatments. Both the perfluorodecalin and the Pluronic F-68 treatments alone and in combination also promoted an increase in biomass, measured as fresh weight gain 30 days after thawing, to a maximum of 38% above control (P < 0.05). These results demonstrate the marked cytoprotectant effects of oxygenated perfluorodecalin and Pluronic F-68, both alone and/or in combination, for plant cells recovered from cryostorage. Such options offer alternative postthaw handling strategies to cells of those plant species which, normally, respond poorly to conventional recovery procedures. Copyright 1997 Academic Press. Copyright 1997Academic Press
研究了向培养基中添加全氟碳化合物以及添加全氟碳化合物与0.01%（w/v）普朗尼克F-68的组合，对粳稻品种台北309悬浮培养细胞冻存后的解冻后活力的有益影响。通过氯化三苯基四氮唑还原法评估，在100毫升玻璃瓶中，解冻后4天的细胞平均活力，经全氟萘烷处理后比对照提高了20%（P<0.05）；在24孔培养皿中进行较小规模复苏时，全氟萘烷处理后的细胞平均活力同样比对照提高了24%（P<0.01）。在另一项评估中，全氟萘烷处理后的解冻后活力比对照处理高出21%（P<0.05）。单独使用0.01%（w/v）普朗尼克F-68处理细胞，解冻后活力也有类似的提高，比对照增加了36%（P<0.05）。当细胞在全氟萘烷和0.01%（w/v）普朗尼克F-68同时存在的情况下复苏时，活力有更显著的协同提高，比对照高出57%（P<0.05）。普朗尼克F-68处理和全氟萘烷处理之间未观察到显著差异。单独或组合使用全氟萘烷和普朗尼克F-68处理，也都促进了生物量的增加，以解冻后30天的鲜重增加量衡量，最高比对照增加了38%（P<0.05）。这些结果表明，全氟萘烷和普朗尼克F-68单独或组合使用，对从冷冻保存中复苏的植物细胞具有显著的细胞保护作用。对于那些通常对传统复苏程序反应不佳的植物物种的细胞，这些方法提供了替代的解冻后处理策略。版权所有1997年学术出版社。版权所有1997年学术出版社
