Post-cryopreservation viability of the benthic freshwater diatom Planothidium frequentissimum depends on light levels

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• 作者：Matthias T. Buhmann ; John G. Day ; Peter G. Kroth
• 关键词：Me2SO ; dimethyl sulfoxide ; CFSE ; carboxy fluorescein succinimidyl ester ; FDA ; fluorescein diacetate ; ROS ; reactive oxygen species
• 刊名：Cryobiology
• 出版年：2013
• 出版时间：August, 2013
• 年：2013
• 卷：67
• 期：1
• 页码：23-29
• 全文大小：914 K

文摘

Over recent years, several planktonic and benthic freshwater diatom taxa have been established as laboratory model strains. In common with most freshwater diatoms the pennate diatom Planothidium frequentissimum suffers irreversible cell shrinkage on prolonged maintenance by serial transfers, without induction of the sexual cycle. Therefore, alternative strategies are required for the long-term maintenance of this strain. Conventional colligative cryopreservation approaches have previously proven unsuccessful with no regrowth. However, in this study using 5 % dimethyl sulfoxide (Me₂SO), controlled cooling at 1 ¡ãC min^?1, automated ice seeding and cooling to ?40 ¡ãC with a final plunge into liquid nitrogen, viability levels were enhanced from 0.3 ¡À 0.4 % to 80 ¡À 3 % , by incorporating a 48 h dark-recovery phase after rewarming. Omission, or reduction, of this recovery step resulted in obvious cell damage with photo-bleaching of pigments, indicative of oxidative-stress induced cell damage, with subsequent deterioration of cellular architecture.