Comparison of cryopreservation methods for the long term storage of the marine diatom Haslea ostrearia (simonsen)

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• 作者：Anaë ; lle Tanniou^a ; ^{anaelle.tanniou@univ-brest.fr} ; Vincent Turpin^b ; ^{vincent.turpin@univ-nantes.fr} ; Thierry Lebeau^c ; ^{thierry.lebeau@univ-nantes.fr}
• 关键词：Cryopreservation ; Immobilization-dehydration ; Diatom ; Me2SO ; MeOH ; Glycerol
• 刊名：Cryobiology
• 出版年：2012
• 期刊代码：166_00112240
• 类别：bio
• 出版时间：August, 2012
• 卷：65
• 期：1
• 页码：45-50
• 文件大小：321 K

摘要

Long term maintenance of microalgal strains by serial subculturing is often expensive and time-consuming. Alternative methods, such as cryopreservation, present several benefits and thus seem more relevant. Our study aimed at comparing two cryopreservation procedures applied to the marine diatom Haslea ostrearia (Simonsen): (1) a two-step freezing method in liquid media using 5%, 10%and 20%MeOH, Me₂SO or Glycerol, and (2) an immobilization-dehydration method consisting in an algal cell entrapped in 0.7 M sucrose dehydrated and air-flow desiccated calcium alginate beads before 鈥渄irect鈥?or 鈥渢wo-step鈥?freezing. Our results showed that the cryopreservation of H. ostrearia was feasible. With the two-step freezing protocol only Me₂SO maintained cell viability without contamination but the low percentage of viability (<10%) prevents its use. Conversely, the immobilization-dehydration methods tested in this study were effective. Average viability of 57%and 77%were obtained with the 鈥渄irect鈥?and the 鈥渢wo step鈥?cooling assays respectively, ensuring preservation of the genetic traits of H. ostrearia.