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Optimized cryopreservation of Ettlia sp. using short cold acclimation and controlled freezing procedure

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Abstract

Despite widespread use as a preservation method, cryopreservation remains recalcitrant for microalgae preservation due to low viability and strain specificity. Herein, a cryopreservation method for Ettlia sp. YC001 was optimized using the following strategies. Cold acclimation at 4 °C for 24 h prior to the freezing procedure enhanced the expression of the cold response pathway and heat-shock proteins, helping to both stabilize the proteins against cold stress and prevent rapid dehydration. Three independent parameters of cryopreservation, the cooling rate, plunge temperature, and thawing temperature, were optimized with various cryoprotectants using a response surface methodology (RSM). Finally, the cellular damage was minimized when using the optimized controlled cooling procedure with an accurate programmable freezer. When applying the optimized conditions, the cell viability of the cryopreserved strain YC001 was > 90% after 1 year. It is believed that the suggested approach can also be applied to other commercial microalgal strains.

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Acknowledgements

This article was written in memory of the dedication, scientific creativity, and warmth of Dr. Hyun-Joon La, who passed away on 13 February 2016.

Funding

This research was supported equally by the Advanced Biomass R&D Center, a Global Frontier Program, funded by the Korean Ministry of Science and ICT (2010-0029723) and the National Research Foundation of Korea (NRF) via a grant funded by the Korean government (NRF-2018R1C1B3009513).

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Authors and Affiliations

  1. Cell Factory Research Center, Korea Research Institute of Bioscience and Biotechnology, 125 Gwahak-ro, Yuseong-gu, Daejeon, 34141, Republic of Korea

    Ji-San Ha, Jun-Woo Lee, Seong-Hyun Seo, Chi-Yong Ahn, Hyung-Gwan Lee & Hee-Mock Oh

  2. Department of Biological Sciences, Sungkyunkwan University, Suwon, 16419, Republic of Korea

    Ji-San Ha

  3. Department of Life Science, Hanyang University, Seoul, 04763, Republic of Korea

    Jun-Woo Lee & Seong-Hyun Seo

  4. Department of Theriogenology and Biotechnology, College of Veterinary Medicine, Gyeongsang National University, Jinju, 52828, Republic of Korea

    Gyu-Jin Rho

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  1. Ji-San Ha
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Correspondence to Hyung-Gwan Lee or Hee-Mock Oh.

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Ha, JS., Lee, JW., Seo, SH. et al. Optimized cryopreservation of Ettlia sp. using short cold acclimation and controlled freezing procedure. J Appl Phycol 31, 2277–2287 (2019). https://doi.org/10.1007/s10811-019-1743-z

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  • DOI: https://doi.org/10.1007/s10811-019-1743-z

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