Lessons from nature for preservation of mammalian cells, tissues, and organs

  • Kelvin G. M. Brockbank
  • Lia H. Campbell
  • Elizabeth D. Greene
  • Matthew C. G. Brockbank
  • John G. Duman
Review
First Online:
Received:
Accepted:

Abstract

The study of mechanisms by which animals tolerate environmental extremes may provide strategies for preservation of living mammalian materials. Animals employ a variety of compounds to enhance their survival, including production of disaccharides, glycerol, and antifreeze compounds. The cryoprotectant glycerol was discovered before its role in amphibian survival. In the last decade, trehalose has made an impact on freezing and drying methods for mammalian cells. Investigation of disaccharides was stimulated by the variety of organisms that tolerate dehydration stress by accumulation of disaccharides. Several methods have been developed for the loading of trehalose into mammalian cells, including inducing membrane lipid-phase transitions, genetically engineered pores, endocytosis, and prolonged cell culture with trehalose. In contrast, the many antifreeze proteins (AFPs) identified in a variety of organisms have had little impact. The first AFPs to be discovered were found in cold water fish; their AFPs have not found a medical application. Insect AFPs function by similar mechanisms, but they are more active and recombinant AFPs may offer the best opportunity for success in medical applications. For example, in contrast to fish AFPs, transgenic organisms expressing insect AFPs exhibit reduced ice nucleation. However, we must remember that nature’s survival strategies may include production of AFPs, antifreeze glycolipids, ice nucleators, polyols, disaccharides, depletion of ice nucleators, and partial desiccation in synchrony with the onset of winter. We anticipate that it is only by combining several natural low temperature survival strategies that the full potential benefits for mammalian cell survival and medical applications can be achieved.

Keywords

Cryopreservation Cryobiology Antifreeze proteins Trehalose Polyols Nucleation Dehydration 
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Notes

Acknowledgements

This work was supported by NSF IOB06-18436 to JGD and R44DK081233 from the National Institute of Diabetes and Digestive and Kidney Diseases to KGMB. The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institute of Diabetes and Digestive and Kidney Diseases or the National Institutes of Health.

References

  1. Amir G.; Horowitz L.; Rubinsky B.; Yousif B. S.; Lavee J.; Smolinskyd A. K. Subzero nonfreezing cryopreservation of rat hearts using antifreeze protein I and antifreeze protein III. Cryobiology 48: 273–282; 2004.PubMedCrossRefGoogle Scholar
  2. Amir G.; Rubinsky B.; Kassif Y.; Horowitz L.; Smolinsky A. K.; Lavee J. Preservation of myocyte structure and mitochondrial integrity in subzero cryopreservation of mammalian hearts for transplantation using antifreeze proteins—an electron microscopy study. Eur J Cardio Thoracic Surg 24: 292–297; 2003.CrossRefGoogle Scholar
  3. Amornwittawat N.; Wang S.; Banatlao J.; Chung M.; Velasco E.; Duman J. G.; Wen X. Effects of polyhydroxy compounds on beetle antifreeze protein activity. Biochimica et Biophysica Acta, Proteins and Proteomics 1794: 341–346; 2009.CrossRefGoogle Scholar
  4. Amornwittawat N.; Wang S.; Duman J. G.; Wen X. Polycarboxylates enhance beetle antifreeze protein activity. Biochim Biophys Acta: Proteins Proteomics 1784: 1942–1948; 2008.CrossRefGoogle Scholar
  5. Andorfer C. A.; Duman J. G. Isolation and characterization of cDNA clones encoding antifreeze proteins of the pyrochroid beetle Dendroides canadensis. J Insect Physiol 46: 365–372; 2000.PubMedCrossRefGoogle Scholar
  6. Arav A.; Ramsbottom G.; Baguis A.; Rubinsky B.; Roche J. F.; Boland M. P. Vitrification of bovine and ovine embryos with the MDS technique and antifreeze proteins. Cryobiology 30: 621–622; 1993.Google Scholar
  7. Beattie G. M.; Crowe J. H.; Lopez A. D.; Cirulli V.; Ricordi C.; Hayek A. Trehalose: a cryoprotectant that enhances recovery and preserves function of human pancreatic islets after long-term storage. Diabetes 46: 519–523; 1997.PubMedCrossRefGoogle Scholar
  8. Bennett V. A.; Sformo T.; Walters K.; Toien O.; Jeannet K.; Hochstrasser R.; Pan Q.; Serianni A. S.; Barnes B. M.; Duman J. G. Comparative overwintering physiology of Alaska and Indiana populations of the beetle Cucujus clavipes (Fabricus): roles of antifreeze proteins, polyols, dehydration, and diapause. J Exp Biol 208: 4467–4477; 2005.PubMedCrossRefGoogle Scholar
  9. Brockbank K. G. M. Essentials of cryobiology. In: Brockbank K. G. M. (ed) Principles of autologous, allogeneic, and cryopreserved venous transplantation. RG Landes Company, Austin, pp 91–102; 1995.Google Scholar
  10. Brockbank K. G. M.; Campbell L. H.; Ratcliff K. M.; Sarver K. A. Method for treatment of cellular materials with sugars prior to preservation. U.S. Patent #7,270,946; 2007.Google Scholar
  11. Brockbank K. G. M.; Chen Z.; Greene E. D., Duman J. G. Anti-freeze proteins enable sub-zero preservation of tissue functions without freezing. Regenerative Medicine: Advancing Next Generation Therapies, abstract. 2009Google Scholar
  12. Brockbank K. G. M.; Taylor M. J. Tissue preservation. In: Baust J. G. (eds) Advances in biopreservation, Chapter 8. CRC Press, Boca Raton, pp 157–196; 2007.Google Scholar
  13. Buchanan S. S.; Menze M. A.; Hand S. C.; Pyatt D. W.; Carpenter J. F. Cell Preserv Technol 3(4): 212–222; 2005.CrossRefGoogle Scholar
  14. Cheng C. C. M.; DeVries A. L. Origins and evolution of fish antifreeze proteins. In: Ewart K. V.; Hew C. L. (eds) Fish antifreeze proteins. World Scientific, New Jersey, pp 83–108; 2002.CrossRefGoogle Scholar
  15. Crowe J. H.; Crowe L. M.; Wolkers W.; Tsvetkova N. M. et al. Stabilization of mammalian cells in the dry state. In: Baust J. G.; Baust J. M. (eds). CRC Press, Taylor and Francis Group, Boca Raton, USA, pp 384–411; 2007Google Scholar
  16. DeVries A. L. Glycoproteins as biological antifreeze agents in Antarctic fishes. Science 172: 1152–1155; 1971.PubMedCrossRefGoogle Scholar
  17. DeVries A. L. Antifreeze glycopeptides and peptides: interactions with ice and water. Meth Enzymol 127: 293–303; 1986.PubMedCrossRefGoogle Scholar
  18. DeVries A. L.; Wohlschlag D. E. Freezing resistance in some Antarctic fishes. Science 163: 1073–1075; 1969.PubMedCrossRefGoogle Scholar
  19. DeVries AL. Ice, antifreeze proteins and antifreeze genes in polar fishes. In: Barnes BM and Carey HV, eds., Life in the Cold: evolution, mechanism, adaptation and application. University of Alaska Press, Fairbanks, pp. 307-316, 2004Google Scholar
  20. Duman J. G. Antifreeze and ice nucleator proteins in terrestrial arthropods. Ann Rev Physiol 63: 327–357; 2001.CrossRefGoogle Scholar
  21. Duman J. G.; Bennett V. A.; Sformo T.; Hochstrasser R.; Barnes B. M. Antifreeze proteins in Alaskan insects and spiders. J Insect Physiol 50: 259–266; 2004.PubMedCrossRefGoogle Scholar
  22. Duman J. G.; Neven L. G.; Beals J. M.; Olsen K. R.; Castellino F. J. Freeze tolerance adaptations, including protein and lipoproten ice nucleators, in larvae of the cranefly Tipula trivittata. J Insect Physiol 31: 1–9; 1985.CrossRefGoogle Scholar
  23. Duman J. G.; Olsen T. M. Thermal hysteresis activity in bacteria, fungi and primitive plants. Cryobiology 30: 322–328; 1993.CrossRefGoogle Scholar
  24. Duman J. G.; Parmalee D.; Goetz F. W.; Li N.; Wu D. W.; Benjamin T. Molecular characterization and sequencing of antifreeze proteins from larvae of the beetle Dendroides canadensis. J Comp Physiol B 168: 225–232; 1998.PubMedCrossRefGoogle Scholar
  25. Duman J. G.; Walters K. R.; Sformo T.; Carrasco M. A.; Nickell P.; Barnes B. M. Antifreeze and ice nucleator proteins. In: Denlinger, D., and Lee, R. E., eds., Low Temperature Biology of Insects. Cambridge University Press, Cambridge, UK, pp. 59-90; 2010.Google Scholar
  26. Eroglu A.; Russo M. J.; Bieganski R.; Fowler A.; Cheley S.; Bayley H.; Toner M. Intracellular trehalose improves the survival of cryopreserved mammalian cells. Nat Biotechnol 18: 163–167; 2000.PubMedCrossRefGoogle Scholar
  27. Eroglu A.; Toner M.; Toth T. L. Beneficial effect of microinjected trehalose on the cryosurvival of human oocytes. Fertil Steril 77(1): 152–158; 2002.PubMedCrossRefGoogle Scholar
  28. Graether S. P.; Sykes B. D. Cold survival in freeze intolerant insects: the structure and function of beta-helical antifreeze proteins. Eur J Biochem 271: 3285–3296; 2004.PubMedCrossRefGoogle Scholar
  29. Griffith M.; Yaish M. W. Antifreeze proteins in plants: a tale of two activities. Trends Plant Sci 9: 399–405; 2004.CrossRefGoogle Scholar
  30. Hansen T. N.; Smith K. M.; Brockbank K. G. M. Type I antifreeze protein attenuates cell recoveries following cryopreservation. Transpl Proc 25(6): 3186–3188; 1993.Google Scholar
  31. Huang T.; Nicodemus J.; Zarka D. G.; Thomashow M. F.; Duman J. G. Expression of an insect (Dendroides canadensis) antifreeze protein in Arabidopsis thaliana results in a decrease in plant freezing temperature. Plant Mol Biol 50: 333–344; 2002.PubMedCrossRefGoogle Scholar
  32. Jia X.; Davies P. L. Antifreeze proteins: an unusual receptor–ligand interaction. Trends Biochem Sci 27: 101–106; 2002.PubMedCrossRefGoogle Scholar
  33. Karow A. M. Biophysical and chemical considerations in cryopreservation. In: Karow A. M.; Pegg D. E. (eds) Organ preservation for transplantation. Marcel Dekker, Inc., New York, pp 113; 1981.Google Scholar
  34. Knight C. A.; Duman J. G. Inhibition of recrystallization of ice by insect thermal hysteresis proteins: a possible cryoprotective role. Cryobiology 23: 256–262; 1986.CrossRefGoogle Scholar
  35. Langeaux D.; Huhtinen M.; Koskinen E.; Palmer E. Effect of antifreeze protein on the cooling and freezing of equine embryos as measured by DAPI-staining. Equine Vet J 25: 85–87; 1997.Google Scholar
  36. Lee C. Y.; Rubinsky B.; Fletcher G. L. Hypothermic preservation of whole mammalian liver with antifreeze proteins. Cryo Lett 13: 59–66; 1992.Google Scholar
  37. Lee R. E. A primer on insect cold tolerance. In: Denlinger D. L., Lee R. E. (eds) Low temperature biology of insects. Cambridge University Press, Cambridge, pp 3–34; 2010.Google Scholar
  38. Li N.; Andorfer C. A.; Duman J. G. Enhancement of insect antifreeze protein activity by low molecular weight solutes. J Exp Biol 201: 2243–2251; 1998.PubMedGoogle Scholar
  39. Lin X.; O’Tousa J. E.; Duman J. G. Expression of two self-enhancing antifreeze proteins from Dendroides canadensis in Drosophila melanogaster. J Insect Physiol 56: 341–349; 2010.PubMedCrossRefGoogle Scholar
  40. Mazur P. Freezing of living cells: mechanisms and implications. Am J Physiol 247(Cell Physiol 16): C125–C142; 1984.PubMedGoogle Scholar
  41. Merryman H. Foreword. In: Fuller B. J.; Lane N.; Benson E. E. (eds) Life in the frozen state. CRC Press, Baton Rouge; 2004.Google Scholar
  42. Miyamoto Y.; Suzuki S.; Nomura K.; Enosawa S. Improvement of hepatocyte viability after cryopreservation by supplementation of long-chain oligosaccharide in the freezing medium in rats and humans. Cell Transplant 15: 911–919; 2006.PubMedCrossRefGoogle Scholar
  43. Mondal B. A simple method for cryopreservation of MDBK cells using trehalose and storage at −80°C. Cell Tissue Bank 10(4): 341–344; 2009.PubMedCrossRefGoogle Scholar
  44. Mugnano J. A.; Wang T.; Layne Jr. J. R.; De Vries A. L.; Lee R. E. Antifreeze glycoproteins promote lethal intracellular freezing of rat cardiomyocytes at high subzero temperatures. Cryobiology 32: 556–557; 1995.Google Scholar
  45. Nicodemus J.; O’Tousa J. E.; Duman J. G. Expression of a beetle, Dendroides canadensis, antifreeze protein in Drosophila melanogaster. J Insect Physiol 52: 888–896; 2006.PubMedCrossRefGoogle Scholar
  46. Norris M. M.; Aksan A.; Sugimachi K.; Toner M. 3-O-methyl-D-glucose improves desiccation tolerance of keratinocytes. Tissue Eng 12: 1873–1879; 2006.PubMedCrossRefGoogle Scholar
  47. Oliver A. E.; Jamil K.; Crowe J. H.; Tablin F. Loading Human Mesenchymal Stem Cells with Trehalose by Fluid-Phase Endocytosis. Cell Preserv Technol 2(1): 35–49; 2004.CrossRefGoogle Scholar
  48. Pegg D. E. Antifreeze proteins. Cryobiology 29: 774–782; 1992.Google Scholar
  49. Petraya N.; Marshall C. B.; Celik Y.; Davies P. L.; Braslovsky I. Direct visualization of spruce budworm antifreeze protein interacting with ice. Basal plane affinity confers hyperactivity. Biophys J 95: 333–341; 2008.CrossRefGoogle Scholar
  50. Polge C.; Smith A. Y.; Parkes A. S. Revival of spermatozoa after vitrification and de-hydration at low temperatures. Nature 164: 666; 1949.PubMedCrossRefGoogle Scholar
  51. Raymond J. A.; DeVries A. L. Adsorption inhibition as a mechanism of freezing resistance in polar fishes. Proceeding Natl Acad Sci USA 74: 2589–2593; 1977.CrossRefGoogle Scholar
  52. Raymond J. A.; Wilson P. W.; DeVries A. L. Inhibition of ice on non-basal planes of ice by fish antifreeze. Proceeding Natl Acad Sci USA 86: 881–885; 1989.CrossRefGoogle Scholar
  53. Redmond J.; Bolin R. B.; Cheney B. A. Glycerol-glucose cryopreservation of platelets. In vivo and in vitro observations. Transfusion 23(3): 213–214; 1983.PubMedCrossRefGoogle Scholar
  54. Rubinsky B.; Arav A.; De Vries A. L. Cryopreservation of oocytes using directional cooling and antifreeze glycoproteins. Cryo Lett 12: 93–106; 1991a.Google Scholar
  55. Rubinsky B.; Arav A.; De Vries A. L. The cryoprotective effect of antifreeze glycopeptides from Antartic fishes. Cryobiology 29: 69–79; 1992.PubMedCrossRefGoogle Scholar
  56. Rubinsky B.; Arav A.; Flecher G. L. Hypothermic protection—a fundamental property of antifreeze proteins. Biochem Biophys Res Commun 180: 566–571; 1991b.PubMedCrossRefGoogle Scholar
  57. Rubinsky B.; Arav A.; Hong J. S.; Lee C. Y. Freezing of mammalian livers with glycerol and antifreeze proteins. Biomech Biophys Res Commun 200(2): 732–741; 1994.CrossRefGoogle Scholar
  58. Sformo T.; Walters K. R.; Jeannette K.; McIntyre J.; Wowk B.; Fahy G.; Barnes B. M.; Duman J. G. Supercooling, vitrification and limited survival to −100°C in larvae of the Alaskan beetle Cucujus clavipes puniceus (Coleoptera: Cucujidae). J Exp Biol 213: 502–509; 2010.PubMedCrossRefGoogle Scholar
  59. Storey K. B.; Storey J. M. Biochemistry of cryoprotectants. In: Denlinger D. L.; Lee R. E. (eds) Insects at low temperature. Chapman and Hall, New York, pp 64–93; 1991.Google Scholar
  60. Storey K. B.; Storey J. M. Physiology, biochemistry, and molecular biology of vertebrate freeze tolerance: the wood frog. In: Fuller B. J.; Lane N.; Benson E. E. (eds) Life in the frozen state. CRC Press, Baton Rouge, pp 243–274; 2004.CrossRefGoogle Scholar
  61. Tonczak M. M., Crowe J. H. The interaction of antifreeze proteins with model membrane and cells. In: Ewart K. V.; Hew C. L. (eds) Fish antifreeze proteins. World Scientific, New Jersey, pp 187–212; 2002Google Scholar
  62. Walters K. R.; Serianni A. S.; Sformo T.; Barnes B. M.; Duman J. G. A thermal hysteresis-producing xylomannan antifreeze in a freeze tolerant Alaskan beetle. Proceeding Natl Acad Sci USA 106: 20210–20215; 2009.CrossRefGoogle Scholar
  63. Wang L.; Duman J. G. Antifreeze proteins of the beetle Dendroides canadensis enhance one another’s activities. Biochemistry 44: 10305–10312; 2005.PubMedCrossRefGoogle Scholar
  64. Wang L.; Duman J. G. A thaumatin-like protein from larvae of the beetle Dendroides canadensis enhances the activity of antifreeze proteins. Biochemistry 45: 1278–1284; 2006.PubMedCrossRefGoogle Scholar
  65. Wang T.; Zhu Q.; Yang X.; Layne Jr. J. R.; DeVries A. L. Antifreeze glycoproteins from the antartic notothenoid fishes fail to protect the rat cardiac explant during hypothermic and freezing preservation. Cryobiology 31: 185–192; 1994.PubMedCrossRefGoogle Scholar
  66. Wharton D. A.; Pow B.; Kristensen M.; Ramlov H.; Marshall C. J. Ice-active proteins and cryoprotectants from the New Zealand alpine cockroach Celatoblatta quinquemaculta. J Insect Physiol 55: 27–31; 2009.PubMedCrossRefGoogle Scholar
  67. Wolkers W. F.; Walker N. J.; Tablin F.; Crowe J. H. Human platelets loaded with trehalose survive freeze-drying. Cryobiology 42: 79–87; 2001.PubMedCrossRefGoogle Scholar
  68. Younis A. I.; Rooks B.; Khan S.; Gould K. B. The effect of antifreeze peptide III and insulin transferrin selenium (ITS) on cryopreservation of chimpanzee spermatozoa. J Androl 19: 207–214; 1998.PubMedGoogle Scholar
  69. Zachariassen K. E.; Hammel H. T. Nucleating agents in the haemolymph of insects tolerant to freezing. Nature 262: 285–287; 1976.PubMedCrossRefGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2010

Authors and Affiliations

  • Kelvin G. M. Brockbank
    • 1
    • 2
  • Lia H. Campbell
    • 1
  • Elizabeth D. Greene
    • 1
  • Matthew C. G. Brockbank
    • 1
  • John G. Duman
    • 3
  1. 1.Cell & Tissue Systems, Inc.North CharlestonUSA
  2. 2.Institute for Bioengineering and BioscienceGeorgia Institute of TechnologyAtlantaUSA
  3. 3.Department of Biological SciencesUniversity of Notre DameNotre DameUSA

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