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Cryopreservative Effects of the Recombinant Ice-Binding Protein from the Arctic Yeast Leucosporidium sp. on Red Blood Cells

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Abstract

Antifreeze proteins (AFPs) have important functions in many freeze-tolerant organisms. The proteins non-colligatively lower the freezing point and functionally inhibit ice recrystallization in frozen solutions. In our previous studies, we found that the Arctic yeast Leucosporidium sp. produces an AFP (LeIBP), and that the protein could be successfully produced in Pichia expression system. The present study showed that recombinant LeIBP possesses the ability to reduce the damage induced to red blood cells (RBCs) by freeze thawing. In addition to 40 % glycerol, both 0.4 and 0.8 mg/ml LeIBPs significantly reduced freeze–thaw-induced hemolysis at either rapid- (45 °C) or slow-warming (22 °C) temperatures. Post-thaw cell counts of the cryopreserved RBCs were dramatically enhanced, in particular, in 0.8 mg/ml LeIBP. Interestingly, the cryopreserved cells in the presence of LeIBP showed preserved cell size distribution. These results indicate that the ability of LeIBP to inhibit ice recrystallization helps the RBCs avoid critically damaging electrolyte concentrations, which are known as solution effects. Considering all these data, LeIBP can be thought of as a key component in improving RBC cryopreservation efficiency.

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Acknowledgments

The authors wish to thank So Ra Yoon for helping in blood collection. This work was supported by grants from Korea Polar Research Institute (PE11100) and Korea Research Council of Fundamental Science and Technology (PG11010 and PG12010).

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Authors and Affiliations

  1. Division of Polar Life Sciences, Korea Polar Research Institute, Incheon, 406-840, Republic of Korea

    Sung Gu Lee, Hye Yeon Koh, Jun Hyuck Lee & Hak Jun Kim

  2. Division of Polar Climate Research, Korea Polar Research Institute, Incheon, 406-840, Republic of Korea

    Sung-Ho Kang

  3. Department of Polar Sciences, University of Science and Technology, Incheon, 406-840, Republic of Korea

    Sung Gu Lee, Jun Hyuck Lee, Sung-Ho Kang & Hak Jun Kim

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  1. Sung Gu Lee
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Correspondence to Hak Jun Kim.

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Sung Gu Lee and Hye Yeon Koh contributed equally to this manuscript.

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Lee, S.G., Koh, H.Y., Lee, J.H. et al. Cryopreservative Effects of the Recombinant Ice-Binding Protein from the Arctic Yeast Leucosporidium sp. on Red Blood Cells. Appl Biochem Biotechnol 167, 824–834 (2012). https://doi.org/10.1007/s12010-012-9739-z

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