impact, status, and future directions. Biol
Reprod 78:2–12
9. Tedder RS, Zuckerman MA, Goldstone AH,
Hawkins AE, Fielding A, Briggs EM,
Irwin D, Blair S, Gorman AM, Patterson KG,
Hawkins AE, Gormon AM, Heptonstall J,
Irwin D (1995) Hepatitis B transmission from
contaminated cryopreservation tank. Lancet
346:137–140
10. Bielanski A, Nadin-Davis S, Sapp T, LutzeWallace C (2000) Viral contamination of
embryos cryopreserved in liquid nitrogen.
Cryobiology 40:110–116
11. Meryman HT, Kafig E (1959) Survival of spermatozoa
following
drying.
Nature
184:470–471
12. Yushchenko NP (1957) Proof of the possibility
of preserving mammalian spermatozoa in a
dried state. Proc Lenin Acad Agric Sci
22:37–40
13. Meryman HT (1960) Drying of living mammalian cells. Ann N Y Acad Sci 85:729–734
14. Larson EV, Graham EF (1976) Freeze-drying
of spermatozoa. Dev Biol Stand 36:343–348
15. Sherman JK (1954) Freezing and freeze-drying
of human spermatozoa. Fertil Steril 5:357–371
16. Bialy G, Smith VR (1957) Freeze-drying of
bovine spermatozoa. J Dairy Sci 40:739–745
17. Saacke RG, Almquist JO (1961) Freeze-drying
of bovine spermatozoa. Nature 192:995–996
18. Nei T, Nagase H (1961) Attempts to freezedry bull spermatozoa. Low Temp Sci B
19:107–115
19. Meryman HT, Kafig E (1963) Freeze-drying of
bovine spermatozoa. J Reprod Fertil 5:87–94
20. Wakayama T, Yanagimachi R (1998) Development of normal mice from oocytes injected
with freeze-dried spermatozoa. Nat Biotechnol
16:639–641
21. Goto K, Kinoshita A, Takuma Y, Ogawa K
(1990) Fertilisation of bovine oocytes by the
injection of immobilised, killed spermatozoa.
Vet Rec 127:517–520
22. Kaneko T, Kimura S, Nakagata N (2007) Offspring derived from oocytes injected with rat
sperm, frozen or freeze-dried without cryoprotection. Theriogenology 68:1017–1021
23. Kaneko T, Serikawa T (2012) Successful longterm preservation of rat sperm by freezedrying. PLoS One 2012:7
24. Muneto T, Horiuchi T (2011) Full-term development of hamster embryos produced by
injecting freeze-dried spermatozoa into
oocytes. J Mamm Ova Res 28:32–39
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 70:1776–1781
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 67:409–415
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 67:1307–1315
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 57:428–432
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 71:1430–1436
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 80:1033–1044
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
142:529–538
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 26:531–536
33. Sanchez-Partida LG, Simerly CR, RamalhoSantos J (2008) Freeze-dried primate sperm
retains early reproductive potential after intracytoplasmic sperm injection. Fertil Steril
89:742–745
34. Kusakabe H, Yanagimachi R, Kamiguchi Y
(2008) Mouse and human spermatozoa can
be freeze-dried without damaging their chromosomes. Hum Reprod 23:233–239
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 97:1067–1073
36. Kusakabe H, Szczygiel MA, Whittingham DG,
Yanagimachi R (2001) Maintenance of genetic
integrity in frozen and freeze-dried mouse
Freeze-Drying of Mammalian Sperm
729
Reprod 78:2–12
9. Tedder RS, Zuckerman MA, Goldstone AH,
Hawkins AE, Fielding A, Briggs EM,
Irwin D, Blair S, Gorman AM, Patterson KG,
Hawkins AE, Gormon AM, Heptonstall J,
Irwin D (1995) Hepatitis B transmission from
contaminated cryopreservation tank. Lancet
346:137–140
10. Bielanski A, Nadin-Davis S, Sapp T, LutzeWallace C (2000) Viral contamination of
embryos cryopreserved in liquid nitrogen.
Cryobiology 40:110–116
11. Meryman HT, Kafig E (1959) Survival of spermatozoa
following
drying.
Nature
184:470–471
12. Yushchenko NP (1957) Proof of the possibility
of preserving mammalian spermatozoa in a
dried state. Proc Lenin Acad Agric Sci
22:37–40
13. Meryman HT (1960) Drying of living mammalian cells. Ann N Y Acad Sci 85:729–734
14. Larson EV, Graham EF (1976) Freeze-drying
of spermatozoa. Dev Biol Stand 36:343–348
15. Sherman JK (1954) Freezing and freeze-drying
of human spermatozoa. Fertil Steril 5:357–371
16. Bialy G, Smith VR (1957) Freeze-drying of
bovine spermatozoa. J Dairy Sci 40:739–745
17. Saacke RG, Almquist JO (1961) Freeze-drying
of bovine spermatozoa. Nature 192:995–996
18. Nei T, Nagase H (1961) Attempts to freezedry bull spermatozoa. Low Temp Sci B
19:107–115
19. Meryman HT, Kafig E (1963) Freeze-drying of
bovine spermatozoa. J Reprod Fertil 5:87–94
20. Wakayama T, Yanagimachi R (1998) Development of normal mice from oocytes injected
with freeze-dried spermatozoa. Nat Biotechnol
16:639–641
21. Goto K, Kinoshita A, Takuma Y, Ogawa K
(1990) Fertilisation of bovine oocytes by the
injection of immobilised, killed spermatozoa.
Vet Rec 127:517–520
22. Kaneko T, Kimura S, Nakagata N (2007) Offspring derived from oocytes injected with rat
sperm, frozen or freeze-dried without cryoprotection. Theriogenology 68:1017–1021
23. Kaneko T, Serikawa T (2012) Successful longterm preservation of rat sperm by freezedrying. PLoS One 2012:7
24. Muneto T, Horiuchi T (2011) Full-term development of hamster embryos produced by
injecting freeze-dried spermatozoa into
oocytes. J Mamm Ova Res 28:32–39
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 70:1776–1781
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 67:409–415
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 67:1307–1315
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 57:428–432
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 71:1430–1436
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 80:1033–1044
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
142:529–538
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 26:531–536
33. Sanchez-Partida LG, Simerly CR, RamalhoSantos J (2008) Freeze-dried primate sperm
retains early reproductive potential after intracytoplasmic sperm injection. Fertil Steril
89:742–745
34. Kusakabe H, Yanagimachi R, Kamiguchi Y
(2008) Mouse and human spermatozoa can
be freeze-dried without damaging their chromosomes. Hum Reprod 23:233–239
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 97:1067–1073
36. Kusakabe H, Szczygiel MA, Whittingham DG,
Yanagimachi R (2001) Maintenance of genetic
integrity in frozen and freeze-dried mouse
Freeze-Drying of Mammalian Sperm
729