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A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen.

Lee HL, Kim SH, Ji DB, Kim YJ - J. Vet. Sci. (2009)

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Obstetrics and Theriogenology, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea.

ABSTRACT
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.

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Related in: MedlinePlus

Sperm plasma-membrane integrity evaluated by carboxyfluoresceindiacetate/propidium iodide (CFDA/PI) fluorescent staining and hypo-osmotic swelling test (HOST) in treated and control samples. Data are presented as mean ± SE. a,b,c,dDifferent superscripts indicate significant differences among treatments within an evaluation method (p < 0.05, n = 6).
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Figure 2: Sperm plasma-membrane integrity evaluated by carboxyfluoresceindiacetate/propidium iodide (CFDA/PI) fluorescent staining and hypo-osmotic swelling test (HOST) in treated and control samples. Data are presented as mean ± SE. a,b,c,dDifferent superscripts indicate significant differences among treatments within an evaluation method (p < 0.05, n = 6).

Mentions: The percentage of intact plasma-membrane was identified by CFDA/PI fluorescent staining and hypo-osmotic swelling test (HOST) (Fig. 2). Spermatozoa obtained by glass wool filtration had the highest percentage of intact membrane from the two evaluation methods (p < 0.05). In CFDA/PI fluorescent staining, the different spermatozoa treatments significantly increased the percentage of spermatozoa with intact plasma-membrane versus control. The percentage of spermatozoa with intact plasma-membrane showed greater value in the order written; control (54.75 ± 8.59%), Sephadex filtration (66.93 ± 6.06%), Percoll separation (74.95 ± 4.43%) and glass wool filtration samples (87.07 ± 1.77%) (p < 0.05). In HOST, the percentages of intact plasma-membrane were higher in glass wool filtration (75.52 ± 3.96%) and Percoll separation samples (58.38 ± 2.22%) than in control samples (44.97 ± 3.54%) (p < 0.05). But Sephadex filtration samples (54.00 ± 5.19%) were not significantly difference with control and Percoll separation samples.


A comparative study of Sephadex, glass wool and Percoll separation techniques on sperm quality and IVF results for cryopreserved bovine semen.

Lee HL, Kim SH, Ji DB, Kim YJ - J. Vet. Sci. (2009)

Sperm plasma-membrane integrity evaluated by carboxyfluoresceindiacetate/propidium iodide (CFDA/PI) fluorescent staining and hypo-osmotic swelling test (HOST) in treated and control samples. Data are presented as mean ± SE. a,b,c,dDifferent superscripts indicate significant differences among treatments within an evaluation method (p < 0.05, n = 6).
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2801134&req=5

Figure 2: Sperm plasma-membrane integrity evaluated by carboxyfluoresceindiacetate/propidium iodide (CFDA/PI) fluorescent staining and hypo-osmotic swelling test (HOST) in treated and control samples. Data are presented as mean ± SE. a,b,c,dDifferent superscripts indicate significant differences among treatments within an evaluation method (p < 0.05, n = 6).
Mentions: The percentage of intact plasma-membrane was identified by CFDA/PI fluorescent staining and hypo-osmotic swelling test (HOST) (Fig. 2). Spermatozoa obtained by glass wool filtration had the highest percentage of intact membrane from the two evaluation methods (p < 0.05). In CFDA/PI fluorescent staining, the different spermatozoa treatments significantly increased the percentage of spermatozoa with intact plasma-membrane versus control. The percentage of spermatozoa with intact plasma-membrane showed greater value in the order written; control (54.75 ± 8.59%), Sephadex filtration (66.93 ± 6.06%), Percoll separation (74.95 ± 4.43%) and glass wool filtration samples (87.07 ± 1.77%) (p < 0.05). In HOST, the percentages of intact plasma-membrane were higher in glass wool filtration (75.52 ± 3.96%) and Percoll separation samples (58.38 ± 2.22%) than in control samples (44.97 ± 3.54%) (p < 0.05). But Sephadex filtration samples (54.00 ± 5.19%) were not significantly difference with control and Percoll separation samples.

Bottom Line: 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.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.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of Veterinary Obstetrics and Theriogenology, College of Veterinary Medicine, Chonbuk National University, Jeonju 561-756, Korea.

ABSTRACT
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.

Show MeSH
Related in: MedlinePlus