A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen.
The aim of this study was to compare the effects of spermatozoa separation techniques on sperm quality and in-vitro fertilization (IVF) results for cryopreserved bovine semen. Sephadex, glass wool and Percoll gradient separation techniques were used for sperm separation and sperm motility, morphology and membrane integrity were evaluated before and after separation. Also, cleavage and blastocyst developmental rate were investigated after IVF with sperm recovered by each separation technique. The motility of samples obtained by the three separation techniques were greater compared to the control samples (p < 0.05). The percentage of spermatozoa with intact plasma-membrane integrity, identified by 6-carboxyfluoresceindiacetate/ propidium iodide fluorescent staining and the hypo-osmotic swelling test, was highest in the glass wool filtration samples (p < 0.05). The cleavage and blastocyst rate of total oocytes produced from glass wool filtration samples were also higher than the control and Sephadex filtration samples (p < 0.05), but were not significantly different from Percoll separation samples. However, a significantly greater number of cleaved embryos produced by glass wool filtration developed to blastocyst stage than those produced by Percoll separation (p < 0.05). These results indicate that spermatozoa with good quality can be achieved by these three separation techniques and can be used for bovine IVF. In particular, it suggests that glass wool filtration would be the most effective method of the three for improving sperm quality and embryo production for cryopreserved bovine spermatozoa.
Lee
Hae-Lee
HL
Laboratory of Veterinary Obstetrics and Theriogenology, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea.
Kim
Sue-Hee
SH
Ji
Dong-Beom
DB
Kim
Yong-Jun
YJ
D000818:Animals; D002417:Cattle; D015925:Cryopreservation / Q000379:methods / Q000662:veterinary*; D003584:Cytological Techniques / Q000379:methods / Q000662:veterinary*; D003637:DEAE-Dextran; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D005898:Glass; D008297:Male; D012662:Semen Preservation / Q000379:methods / Q000662:veterinary*; D013094:Spermatozoa / Q000502:physiology*; D015053:Zygote / Q000166:cytology
D003637:DEAE-Dextran
J Androl. 2006 Mar-Apr;27(2):176-88
16304212
Fertil Steril. 1996 Sep;66(3):459-62
8751748
Theriogenology. 2000 Dec 1;54(9):1343-57
11191860
Theriogenology. 1995 Jan 15;43(2):439-49
16727636
Hum Reprod. 1997 Mar;12(3):509-13
9130752
Hum Reprod. 1993 Nov;8(11):1856-62
8288751
J Dairy Sci. 1990 Jan;73(1):91-7
2312886
Andrologia. 2001 May;33(3):143-50
11380329
Theriogenology. 1999 Mar;51(4):721-7
10728997
Hum Reprod. 1999 Aug;14(8):2015-9
10438419
Hum Reprod. 1993 Sep;8(9):1438-42
8253932
J Androl. 1991 May-Jun;12(3):201-8
1655686
J Androl. 2002 Jul-Aug;23(4):498-502
12065456
Arch Androl. 1994 Mar-Apr;32(2):157-62
8166579
Arch Androl. 1994 Jan-Feb;32(1):5-11
8122936
Biol Reprod. 1975 Mar;12(2):260-74
1122333
Gamete Res. 1988 Jun;20(2):159-64
3235033
Fertil Steril. 1997 Sep;68(3):506-9
9314923
J Dairy Sci. 1972 May;55(5):614-20
4623377
Free Radic Biol Med. 2000 Feb 15;28(4):529-36
10719234
Theriogenology. 1995 Oct 15;44(6):859-69
16727781
Cytogenet Cell Genet. 2000;90(3-4):219-26
11124518
Theriogenology. 1996 Sep;46(4):679-86
16727933
J Reprod Fertil. 1988 Nov;84(2):551-6
3199373
Fertil Steril. 1989 Apr;51(4):685-90
2924937
Cryobiology. 1982 Feb;19(1):20-8
7067484
J Endocrinol Invest. 2000 Nov;23(10):697-710
11097436
Hum Fertil (Camb). 2000;3(1):26-30
11844351
J Androl. 2001 Nov-Dec;22(6):927-36
11700855
Andrologia. 2001 Jul;33(4):223-30
11472334
Hum Reprod. 1988 Jan;3(1):85-8
3350941
Theriogenology. 2005 Jan 1;63(1):160-78
15589282
Biol Reprod. 1991 Feb;44(2):256-60
2009327
J Androl. 1990 Jan-Feb;11(1):73-88
2179184
Andrologia. 2002 Dec;34(6):397-403
12472625
Theriogenology. 1996 Jan 15;45(2):513-20
16727814
Theriogenology. 2000 Apr 15;53(7):1477-88
10898217
Andrologia. 1994 Jan-Feb;26(1):33-4
8185058
Fertil Steril. 1977 Feb;28(2):178-81
576215
Theriogenology. 2006 Apr 15;65(7):1360-75
16256187
J Assist Reprod Genet. 1994 Sep;11(8):395-400
7606151
J Assist Reprod Genet. 2000 Jul;17(6):307-9
11042825
Theriogenology. 1997 Mar;47(4):845-56
16728034
Theriogenology. 1994 Aug 1;42(2):351-60
16727543