Separation of Cryopreserved Human Semen Using Sephadex Columns, Washing, or Percoll Gradients

DROBNIS, ERMA Z.; ZHONG, CHANG Q.; OVERSTREET, JAMES W.

doi:10.1002/j.1939-4640.1991.tb00250.x

Cited by 66 publications

References 59 publications

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Semen preparation techniques for intrauterine insemination

Boomsma

Heineman²,

Cohlen

et al. 2004

Cochrane Database of Systematic Reviews

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There is insufficient evidence to recommend any specific preparation technique. Large high quality randomised controlled trials, comparing the effectiveness of a gradient and/ or a swim-up and/ or wash and centrifugation technique on clinical outcome are lacking. Further randomised trials are warranted. Results from studies comparing semen parameters may suggest a preference for gradient technique, but firm conclusions cannot be drawn and the limitations should be taken into consideration.

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Semen preparation techniques for intrauterine insemination

Boomsma

Heineman²,

Cohlen

et al. 2004

Cochrane Database of Systematic Reviews

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Evidence for Increased Lipid Peroxidative Damage and Loss of Superoxide Dismutase Activity as a Mode of Sublethal Cryodamage to Human Sperm During Cryopreservation

ALVAREZ,

STOREY

1992

Journal of Andrology

285

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Cryopreservation of human sperm, now generally required in donor insemination programs, adversely affects the sperm in terms of standard sperm evaluation parameters and fertilizing ability. The freeze‐thaw process appears to produce sublethal damage that appears only after a delay. The authors hypothesized that cryopreservation enhanced peroxidation of sperm membrane lipids, based on previous studies of sperm lipid peroxidation, which showed that the effects of peroxidative damage became evident only after a delay, depending on the peroxidation rate. The effect of cryopreservation on the phospholipid content, the composition of the acyl moieties of the phospholipids, and the activities of the peroxidation protective enzymes, superoxide dimutase (SOD) and glutathione peroxidase plus reductase, in human sperm were examined to test the hypothesis. Parallel determinations were made of the percent motility, the average path velocity of the motile cells, and the time to loss of motility under specified aerobic incubation conditions, which gives a good estimate of the lipid peroxidation rate. The phospholipid content decreases after cryopreservation, with loss of phosphatidylcholine and phosphatidylethanolamine being the more pronounced. Polyunsaturated acyl moieties were also preferentially lost. This loss pattern is observed also from lipid peroxidation. The activities of glutathione peroxidase plus reductase remained unchanged. The sperm SOD activities varied widely between samples before cryopreservation. In all samples there was a decline in SOD activity after freeze—thaw, but the extent of the decline was also widely variable. The time to loss of motility declined in parallel with SOD activity, and a strong correlation (R2 > 0.9) between SOD activity and time to loss of motility was found for all samples, before and after freeze—thaw. The authors conclude that cryopreservation does enhance lipid peroxidation in human sperm, as hypothesized, and that this enhancement is mediated at least in part by the loss of SOD activity occurring during the process.

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Evidence that Membrane Stress Contributes More than Lipid Peroxidation to Sublethal Cryodamage in Cryopreserved Human Sperm: Glycerol and Other Polyols as Sole Cryoprotectant

ALVAREZ,

STOREY

1993

Journal of Andrology

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One effect of cryopreservation on human sperm is sublethal cryodamage, in which cell viability post‐thaw is lost more rapidly at later times than in fresh cells. We hypothesized two modes of sublethal cryodamage: one is peroxidation‐related involving plasma membrane damage due to lipid peroxidation; the other is membrane stress‐related involving membrane embrittlement during phase transitions occurring during freeze‐thaw. If the peroxidation‐related mode contributed substantially to sublethal cryodamage, the hypothesis predicts that lipid peroxidation inhibitors should reduce this damage. To test this prediction, we examined the effect of the lipid peroxidation inhibitors, hypotaurine, bovine serum albumin (BSA), and α‐tocopheroi (Vit. E) on the time to loss of motility (TLM), taken as a measure of cell viability over time, for sperm samples cryopreserved in glycerol plus egg yolk medium. These agents had no effect on TLM of these samples, indicating that this mode contributes little to sublethal cryodamage. If the membrane stress‐related mode contributed, the hypothesis predicts rapid recovery of motility in the presence of egg yolk plus glycerol, but slow recovery in the presence of glycerol alone. It also predicts that an appropriate polyol may be both necessary and sufficient for cryopreservation. In the presence of egg yolk plus glycerol, motility recovery was complete within 5 minutes, but the percent motile cells then decreased linearly with time. With glycerol alone in the range 3–12%, at 5 minutes post‐thaw the percent motile cells was 5–10%, but by 40 minutes post‐thaw had risen to 60–80%, approaching that in the fresh sample, and was maintained up to 4 hours. In the absence of glycerol, the percentage of motile cells post‐thaw was nil and remained nil up to 4 hours. The polyols, erythritol, ribitol, and sorbitol had similar effects to that of glycerol, but the recovery of motility was not as complete. These results indicate that the membrane stress‐related mode contributes substantially to sublethal cryodamage. They also indicate that glycerol and other polyols can function alone as cryoprotectants, but that recovery of motility is slow in these systems.

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Separation of Cryopreserved Human Semen Using Sephadex Columns, Washing, or Percoll Gradients

Cited by 66 publications

References 59 publications

Semen preparation techniques for intrauterine insemination

Semen preparation techniques for intrauterine insemination

Evidence for Increased Lipid Peroxidative Damage and Loss of Superoxide Dismutase Activity as a Mode of Sublethal Cryodamage to Human Sperm During Cryopreservation

Evidence that Membrane Stress Contributes More than Lipid Peroxidation to Sublethal Cryodamage in Cryopreserved Human Sperm: Glycerol and Other Polyols as Sole Cryoprotectant

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