Advertisement

Freeze-Drying of Mammalian Sperm

  • Levent Keskintepe
  • Ali ErogluEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1257)

Abstract

Long-term preservation of mammalian sperm at suprazero temperatures is desired to save storage and space costs as well as to facilitate transport of preserved samples. This can be accomplished by the freeze-drying of sperm samples. Although freeze-drying results in immotile and membrane-compromised sperm, intracytoplasmic sperm injection (ICSI) can be used to introduce such an immotile sperm into an oocyte and thus start the fertilization process. So far, it has been shown that improved freeze-drying protocols preserve chromosomal integrity and oocyte-activating factor(s) at 4 °C for several years and at ambient temperature for approximately 1 month, which permits shipping freeze-dried samples at ambient temperature. This chapter concisely reviews freeze-drying of mammalian sperm first and then presents a simple freeze-drying protocol.

Key words

Sperm Spermatozoa Freezing Freeze-drying Cryopreservation Long-term storage Intracytoplasmic sperm injection ICSI Cryoprotectant Trehalose 

References

  1. 1.
    Walters EM, Benson JD, Woods EJ, Critser JK (2009) The history of sperm cryopreservation. In: Pacey AA, Tomlinson MJ (eds) Sperm banking: theory and practice. Cambridge University Press, Cambridge, UK, pp 1–17Google Scholar
  2. 2.
    Polge C, Smith AU, Parkes AS (1949) Revival of spermatozoa after vitrification and dehydration at low temperatures. Nature 164:666CrossRefGoogle Scholar
  3. 3.
    Polge C (1952) Fertilizing capacity of bull spermatozoa after freezing at 79 degrees C. Nature 169:626–627CrossRefGoogle Scholar
  4. 4.
    Stewart DL (1951) Storage of bull spermatozoa at low temperatures. Vet Rec 63:65–66Google Scholar
  5. 5.
    Bunge RG, Sherman JK (1953) Fertilizing capacity of frozen human spermatozoa. Nature 172:767–768CrossRefGoogle Scholar
  6. 6.
    Sherman JK (1990) Cryopreservation of human semen. In: Keel BA, Webster BW (eds) Handbook of the laboratory diagnosis and treatment of infertility. CRC, Boston, MA, pp 229–260Google Scholar
  7. 7.
    Curry MR (2000) Cryopreservation of semen from domestic livestock. Rev Reprod 5:46–52CrossRefGoogle Scholar
  8. 8.
    Mazur P, Leibo SP, Seidel GE (2008) Cryopreservation of the germplasm of animals used in biological and medical research: importance, impact, status, and future directions. Biol Reprod 78:2–12CrossRefGoogle Scholar
  9. 9.
    Tedder RS, Zuckerman MA, Goldstone AH, Hawkins AE, Fielding A, Briggs EM, Irwin D, Blair S, Gorman AM, Patterson KG et al (1995) Hepatitis B transmission from contaminated cryopreservation tank. Lancet 346:137–140CrossRefGoogle Scholar
  10. 10.
    Bielanski A, Nadin-Davis S, Sapp T, Lutze-Wallace C (2000) Viral contamination of embryos cryopreserved in liquid nitrogen. Cryobiology 40:110–116CrossRefGoogle Scholar
  11. 11.
    Meryman HT, Kafig E (1989) Survival of spermatozoa following drying. Nature 184:470–471CrossRefGoogle Scholar
  12. 12.
    Yushchenko NP (1957) Proof of the possibility of preserving mammalian spermatozoa in a dried state. Proc Lenin Acad Agr Sci 22:37–40Google Scholar
  13. 13.
    Meryman HT (1960) Drying of living mammalian cells. Ann N Y Acad Sci 85:729–734CrossRefGoogle Scholar
  14. 14.
    Larson EV, Graham EF (1976) Freeze-drying of spermatozoa. Dev Biol Stand 36:343–348Google Scholar
  15. 15.
    Sherman JK (1954) Freezing and freeze-drying of human spermatozoa. Fertil Steril 5:357–371Google Scholar
  16. 16.
    Bialy G, Smith VR (1957) Freeze-drying of bovine spermatozoa. J Dairy Sci 40:739–745CrossRefGoogle Scholar
  17. 17.
    Saacke RG, Almquist JO (1961) Freeze-drying of bovine spermatozoa. Nature 192:995–996CrossRefGoogle Scholar
  18. 18.
    Nei T, Nagase H (1961) Attempts to freeze-dry bull spermatozoa. Low Temp Sci Ser B 19:107–115Google Scholar
  19. 19.
    Meryman HT, Kafig E (1963) Freeze-drying of bovine spermatozoa. J Reprod Fertil 5:87–94CrossRefGoogle Scholar
  20. 20.
    Wakayama T, Yanagimachi R (1998) Development of normal mice from oocytes injected with freeze-dried spermatozoa. Nat Biotechnol 16:639–641CrossRefGoogle Scholar
  21. 21.
    Goto K, Kinoshita A, Takuma Y, Ogawa K (1990) Fertilisation of bovine oocytes by the injection of immobilised, killed spermatozoa. Vet Rec 127:517–520Google Scholar
  22. 22.
    Kaneko T, Kimura S, Nakagata N (2007) Offspring derived from oocytes injected with rat sperm, frozen or freeze-dried without cryoprotection. Theriogenology 68:1017–1021CrossRefGoogle Scholar
  23. 23.
    Kaneko T, Serikawa T (2012) Successful long-term preservation of rat sperm by freeze-drying. PLoS One 7:e35043CrossRefGoogle Scholar
  24. 24.
    Muneto T, Horiuchi T (2011) Full-term development of hamster embryos produced by injecting freeze-dried spermatozoa into oocytes. J Mamm Ova Res 28:32–39CrossRefGoogle Scholar
  25. 25.
    Liu JL, Kusakabe H, Chang CC, Suzuki H, Schmidt DW, Julian M, Pfeffer R, Bormann CL, Tian XC, Yanagimachi R, Yang XZ (2004) Freeze-dried sperm fertilization leads to full-term development in rabbits. Biol Reprod 70:1776–1781CrossRefGoogle Scholar
  26. 26.
    Keskintepe L, Pacholczyk G, Machnicka A, Norris K, Curuk MA, Khan I, Brackett BG (2002) Bovine blastocyst development from oocytes injected with freeze-dried spermatozoa. Biol Reprod 67:409–415CrossRefGoogle Scholar
  27. 27.
    Martins CF, Bao SN, Dode MN, Correa GA, Rumpf R (2007) Effects of freeze-drying on cytology, ultrastructure, DNA fragmentation, and fertilizing ability of bovine sperm. Theriogenology 67:1307–1315CrossRefGoogle Scholar
  28. 28.
    Hara H, Abdalla H, Morita H, Kuwayama M, Hirabayashi M, Hochi S (2011) Procedure for bovine ICSI, not sperm freeze-drying, impairs the function of the microtubule-organizing center. J Reprod Dev 57:428–432CrossRefGoogle Scholar
  29. 29.
    Kwon IK, Park KE, Niwa K (2004) Activation, pronuclear formation, and development in vitro of pig oocytes following intracytoplasmic injection of freeze-dried spermatozoa. Biol Reprod 71:1430–1436CrossRefGoogle Scholar
  30. 30.
    Men NT, Kikuchi K, Nakai M, Fukuda A, Tanihara F, Noguchi J, Kaneko H, Linh NV, Nguyen BX, Nagai T, Tajima A (2013) Effect of trehalose on DNA integrity of freeze-dried boar sperm, fertilization, and embryo development after intracytoplasmic sperm injection. Theriogenology 80:1033–1044CrossRefGoogle Scholar
  31. 31.
    Choi YH, Varner DD, Love CC, Hartman DL, Hinrichs K (2011) Production of live foals via intracytoplasmic injection of lyophilized sperm and sperm extract in the horse. Reproduction 142:529–538CrossRefGoogle Scholar
  32. 32.
    Watanabe H, Asano T, Abe Y, Fukui Y, Suzuki H (2009) Pronuclear formation of freeze-dried canine spermatozoa microinjected into mouse oocytes. J Assist Reprod Genet 26:531–536CrossRefGoogle Scholar
  33. 33.
    Sanchez-Partida LG, Simerly CR, Ramalho-Santos J (2008) Freeze-dried primate sperm retains early reproductive potential after intracytoplasmic sperm injection. Fertil Steril 89:742–745CrossRefGoogle Scholar
  34. 34.
    Kusakabe H, Yanagimachi R, Kamiguchi Y (2008) Mouse and human spermatozoa can be freeze-dried without damaging their chromosomes. Hum Reprod 23:233–239CrossRefGoogle Scholar
  35. 35.
    Gianaroli L, Magli MC, Stanghellini I, Crippa A, Crivello AM, Pescatori ES, Ferraretti AP (2012) DNA integrity is maintained after freeze-drying of human spermatozoa. Fertil Steril 97:1067–1073CrossRefGoogle Scholar
  36. 36.
    Kusakabe H, Szczygiel MA, Whittingham DG, Yanagimachi R (2001) Maintenance of genetic integrity in frozen and freeze-dried mouse spermatozoa. Proc Natl Acad Sci U S A 98:13501–13506CrossRefGoogle Scholar
  37. 37.
    Kaneko T, Nakagata N (2006) Improvement in the long-term stability of freeze-dried mouse spermatozoa by adding of a chelating agent. Cryobiology 53:279–282CrossRefGoogle Scholar
  38. 38.
    Nakai M, Kashiwazaki N, Takizawa A, Maedomari N, Ozawa M, Noguchi J, Kaneko H, Shino M, Kikuchi K (2007) Effects of chelating agents during freeze-drying of boar spermatozoa on DNA fragmentation and on developmental ability in vitro and in vivo after intracytoplasmic sperm head injection. Zygote 15:15–24CrossRefGoogle Scholar
  39. 39.
    Kaneko T, Whittingham DG, Yanagimachi R (2003) Effect of pH value of freeze-drying solution on the chromosome integrity and developmental ability of mouse spermatozoa. Biol Reprod 68:136–139CrossRefGoogle Scholar
  40. 40.
    Kaneko T, Whittingham DG, Overstreet JW, Yanagimachi R (2003) Tolerance of the mouse sperm nuclei to freeze-drying depends on their disulfide status. Biol Reprod 69:1859–1862CrossRefGoogle Scholar
  41. 41.
    Kawase Y, Araya H, Kamada N, Jishage K, Suzuki H (2005) Possibility of long-term preservation of freeze-dried mouse spermatozoa. Biol Reprod 72:568–573CrossRefGoogle Scholar
  42. 42.
    Kaneko T, Nakagata N (2005) Relation between storage temperature and fertilizing ability of freeze-dried mouse spermatozoa. Comp Med 55:140–144Google Scholar
  43. 43.
    Martins CF, Dode MN, Bao SN, Rumpf R (2007) The use of the acridine orange test and the TUNEL assay to assess the integrity of freeze-dried bovine spermatozoa DNA. Genet Mol Res 6:94–104Google Scholar
  44. 44.
    Donnelly ET, McClure N, Lewis SE (2001) Cryopreservation of human semen and prepared sperm: effects on motility parameters and DNA integrity. Fertil Steril 76:892–900CrossRefGoogle Scholar
  45. 45.
    Ward MA, Kaneko T, Kusakabe H, Biggers JD, Whittingham DG, Yanagimachi R (2003) Long-term preservation of mouse spermatozoa after freeze-drying and freezing without cryoprotection. Biol Reprod 69:2100–2108CrossRefGoogle Scholar
  46. 46.
    Kawase Y, Tachibe T, Jishage K, Suzuki H (2007) Transportation of freeze-dried mouse spermatozoa under different preservation conditions. J Reprod Dev 53:1169–1174CrossRefGoogle Scholar
  47. 47.
    Kaneko T, Serikawa T (2012) Long-term preservation of freeze-dried mouse spermatozoa. Cryobiology 64:211–214CrossRefGoogle Scholar
  48. 48.
    Hochi S, Watanabe K, Kato M, Hirabayashi M (2008) Live rats resulting from injection of oocytes with spermatozoa freeze-dried and stored for one year. Mol Reprod Dev 75:890–894CrossRefGoogle Scholar
  49. 49.
    Kaneko T, Kimura S, Nakagata N (2009) Importance of primary culture conditions for the development of rat ICSI embryos and long-term preservation of freeze-dried sperm. Cryobiology 58:293–297CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  1. 1.School of MedicineSher Institute for Reproductive Medicine and University of NevadaLas VegasUSA
  2. 2.Department of Neuroscience and Regenerative Medicine; Department of MedicineMedical College of Georgia, Georgia Regents UniversityAugustaUSA
  3. 3.Department of Obstetrics and GynecologyMedical College of Georgia, Georgia Regents UniversityAugustaUSA

Personalised recommendations