title: Short-term storage of human spermatozoa in electrolyte-free medium without freezing maintains sperm chromatin integrity better than cryopreservation.
abstract: Previous attempts to maintain human spermatozoa without freezing were based on short-term storage in component-rich medium and led to fast decline in motility and increased incidence of chromosome breaks. Here we report a new method in which sperm are maintained without freezing in an electrolyte-free medium (EFM) composed of glucose and bovine serum albumin. Human sperm were stored in EFM or human tubal fluid medium (HTFM) or were cryopreserved, and their motility, viability, and DNA integrity were examined at different intervals. Cryopreservation led to significant decline in sperm motility and viability and induced DNA fragmentation. Sperm stored in EFM maintained motility and viability for up to 4 and 7 wk, respectively, much longer than sperm stored in HTFM (<2 and <4 wk, respectively). DNA integrity, assessed with comet assay, was also maintained significantly better in EFM than in HTFM. One-week storage in EFM yielded motility and viability similar to that of cryopreserved sperm, but DNA integrity was significantly higher, resembling that of fresh sperm. After several weeks of storage in EFM, sperm were able to activate oocytes, undergo chromatin remodeling, and form normal zygotic chromosomes after intracytoplasmic sperm injection. This study demonstrated that human spermatozoa can be stored in EFM without freezing for several weeks while maintaining motility, viability, and chromatin integrity and that 1-wk storage in EFM offers better protection of sperm DNA integrity than cryopreservation. Sperm storage in EFM may become a viable option for the physicians working in assisted reproduction technology clinics, which would avoid cryodamage.
authors: Institute for Biogenesis Research, John A. Burns School of Medicine, University of Hawaii, Honolulu, Hawaii, USA.
authors: Jonathan M
authors: JM
authors: Riel
authors: Yasuhiro
authors: Y
authors: Yamauchi
authors: Thomas T F
authors: TT
authors: Huang
authors: John
authors: J
authors: Grove
authors: Monika A
authors: MA
authors: Ward
references: Hum Reprod. 1991 May;6(5):703-8
references: 1939553
references: Hum Reprod. 2010 Apr;25(4):824-38
references: 20139429
references: Hum Reprod. 2010 Oct;25(10):2415-26
references: 20716559
references: Biol Reprod. 1991 Apr;44(4):569-74
references: 2043729
references: Int J Androl. 1999 Jun;22(3):155-62
references: 10367235
references: Mol Hum Reprod. 2010 Jan;16(1):3-13
references: 19648152
references: Zygote. 2004 May;12(2):111-6
references: 15460105
references: Zygote. 2010 Aug;18(3):245-56
references: 20109266
references: Biol Reprod. 2005 Oct;73(4):627-33
references: 15930320
references: Biol Reprod. 2003 May;68(5):1779-86
references: 12606475
references: J Biol Chem. 2008 Jan 4;283(1):1-5
references: 17999957
references: Arch Androl. 2007 Sep-Oct;53(5):275-84
references: 18309900
references: Mol Reprod Dev. 1990 Jun;26(2):101-4
references: 2372392
references: Fertil Steril. 2010 Jan;93(1):159-66
references: 19027111
references: Reprod Biomed Online. 2010 Oct;21(4):456-62
references: 20800544
references: Biol Reprod. 2008 Apr;78(4):761-72
references: 18199884
references: Fertil Steril. 1997 Jan;67(1):142-7
references: 8986699
references: Andrologia. 1984 Sep-Oct;16(5):429-33
references: 6496961
references: Fertil Steril. 2010 Mar 1;93(4):1027-36
references: 20080235
references: Reprod Fertil Dev. 2004;16(5):547-54
references: 15355763
references: Di Yi Jun Yi Da Xue Xue Bao. 2002 Oct;22(10):928-30
references: 12377623
references: Fertil Steril. 2009 Dec;92(6):2109-12
references: 19608173
references: J Androl. 2003 Mar-Apr;24(2):253-62
references: 12634313
references: Biol Reprod. 2007 Oct;77(4):636-47
references: 17596560
references: Int J Androl. 2009 Feb;32(1):46-56
references: 19076252
references: Hum Reprod. 2001 Jun;16(6):1191-9
references: 11387291
references: Genome Biol. 2010;11(6):R66
references: 20573212
references: Hum Reprod. 1993 Apr;8(4):581-6
references: 8501190
references: Biol Reprod. 1998 Nov;59(5):1037-46
references: 9780307
references: Fertil Steril. 2000 Sep;74(3):454-60
references: 10973637
references: Biol Reprod. 2007 Nov;77(5):803-12
references: 17652664
references: Gamete Res. 1989 May;23(1):77-90
references: 2545584
references: Ginekol Pol. 2003 Feb;74(2):108-15
references: 12715419
references: J Androl. 2002 Mar-Apr;23(2):194-201
references: 11868812
references: Arch Androl. 2003 Jan-Feb;49(1):49-55
references: 12647778
references: Fertil Steril. 1997 Aug;68(2):373-5
references: 9240274
references: J Reprod Fertil. 1989 Jul;86(2):679-88
references: 2760894
references: Mol Hum Reprod. 2010 Jan;16(1):48-56
references: 19897543
references: Hum Reprod. 2007 Jan;22(1):174-9
references: 16921163
references: Fertil Steril. 1996 Jun;65(6):1210-3
references: 8641499
references: Hum Reprod. 1996 Sep;11(9):1942-6
references: 8921068
references: Hum Reprod. 2008 Dec;23(12):2663-8
references: 18757447
references: Hum Reprod. 1995 May;10(5):1130-5
references: 7657752
references: Fertil Steril. 1983 Apr;39(4):536-41
references: 6682052
references: Hum Reprod. 1996 Oct;11(10):2170-5
references: 8943524
references: Biol Reprod. 2000 Jul;63(1):341-6
references: 10859277
references: Hum Reprod. 1999 Apr;14(4):1039-49
references: 10221239
references: Cell Tissue Bank. 2000;1(2):133-42
references: 15256959
references: Reproduction. 2006 Jun;131(6):977-88
references: 16735537
references: J Assist Reprod Genet. 1998 Mar;15(3):154-7
references: 9547692
references: Fertil Steril. 2006 Sep;86(3):597-600
references: 16952507
references: J Cell Biochem. 2008 Apr 1;103(5):1636-45
references: 17879959
references: Fertil Steril. 1981 Feb;35(2):205-8
references: 7202744
references: Mol Cell Endocrinol. 2000 Nov 27;169(1-2):3-10
references: 11155950
references: Biol Reprod. 1995 Apr;52(4):709-20
references: 7779992
references: Hum Reprod. 2009 Sep;24(9):2061-70
references: 19525298
references: Hum Mol Genet. 2008 Jul 1;17(13):1922-37
references: 18353795
references: Asian J Androl. 2011 Jan;13(1):36-42
references: 20802502
references: Hum Reprod. 2002 Apr;17(4):990-8
references: 11925396
references: Hum Reprod. 2010 Jul;25(7):1594-608
references: 20447937
references: DNA Cell Biol. 1997 Sep;16(9):1087-97
references: 9324311
references: Fertil Steril. 2008 Jun;89(6):1723-7
references: 17953963
references: Andrologia. 1999 Jul;31(4):211-5
references: 10470411