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

Abstract : 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 "direct" or "two-step" 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 "direct" and the "two step" cooling assays respectively, ensuring preservation of the genetic traits of H. ostrearia.
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Submitted on : Friday, July 27, 2012 - 4:07:50 PM
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Anaelle Tanniou, Vincent Turpin, Thierry Lebeau. Comparison of cryopreservation methods for the long term storage of the marine diatom Haslea ostrearia (simonsen). Cryobiology, Elsevier, 2012, 65 (1), pp.45-50. ⟨10.1016/j.cryobiol.2012.03.011⟩. ⟨hal-00721494⟩

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