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Medium composition for effective slow freezing of embryonic cell lines derived from marine medaka (Oryzias dancena)

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Abstract

This study was conducted to identify optimal medium composition for freezing Oryzias dancena embryonic cell lines. Different freezing media consisting of various concentration of dimethyl sulfoxide (DMSO), fetal bovine serum (FBS), and trehalose were prepared and long-term cultured embryonic cell line was frozen in each freezing medium by conventional slow freezing program for 7 days. Through measurement of viability and growth of post-thaw cells frozen in each freezing medium, it was determined that optimal composition of three components was 10 % DMSO, 20 % FBS, and 0.1 M trehalose. The post-thaw cells frozen in optimal freezing medium showed similar morphology and growth rate with non-frozen cells. Next, this condition was applied to two different sets of experiment; (1) freezing of the same cells during expanded period (57 days) and (2) freezing of short-term cultured cells from other batches for 7 days. The viability of post-thaw cells was significantly low and comparable in set 1 and 2, respectively, when compared with the result of long term-cultured cells frozen in optimal freezing medium for 7 days and similar morphology and growth rate with non-frozen counterparts were detected in the post-thaw cells from both sets. In conclusion, this study first reports the optimal medium composition for freezing O. dancena embryonic cells, which can contribute to fish species preservation as well as improvement of cell-based biotechnology by providing stable cell storage.

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Acknowledgments

This research was supported by Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (NRF-2012R1A1A1011572).

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Authors and Affiliations

  1. Department of Fisheries Biology, Pukyong National University, Busan, 608-737, Korea

    Min Sung Kim & Seung Pyo Gong

  2. Department of Marine Biomaterials and Aquaculture, Pukyong National University, Busan, 608-737, Korea

    Min Sung Kim & Seung Pyo Gong

  3. Department of Animal Biotechnology, Kangwon National University, Chuncheon, 200-701, Korea

    Seung Tae Lee

  4. Major in Biomodulation and Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea

    Jeong Mook Lim

  5. Laboratory of Cell Biotechnology, Department of Marine Biomaterials and Aquaculture, College of Fisheries Science, Pukyong National University, Busan, 608-737, Korea

    Seung Pyo Gong

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  1. Min Sung Kim
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Correspondence to Seung Pyo Gong.

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Kim, M.S., Lee, S.T., Lim, J.M. et al. Medium composition for effective slow freezing of embryonic cell lines derived from marine medaka (Oryzias dancena). Cytotechnology 68, 9–17 (2016). https://doi.org/10.1007/s10616-014-9749-5

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