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Human Reproduction, Vol. 8, No. 5, pp. 717-721, 1993
© 1993 European Society of Human Reproduction and Embryology

other

Human spermatozoal tail hypo-osmotic swelling test, motility characteristics in hypotonic saline, and survival of spermatozoa after cryopreservation

S.Y.W. Chan1,4, A. Pearlstone1, M. Uhler1, M. Tucker3, R. Greenspoon1, A. Leung1,2 and C. Wang1,2

1Divisions of Reproductive Endocrinology and Infertility, Cedars-Sinai Medical Center Suite 1101, 444 South San Vicente Boulevard, Los Angeles, CA 90048 2Endocrinology/Metabolism, Cedars-Sinai Medical Center Suite 1101, 444 South San Vicente Boulevard, Los Angeles, CA 90048 3In Vitro Fertilization Program, Reproductive Biology Associates Atlanta, GA, USA

Correspondence: 4To whom correspondence should be addressed

Normozoospermic semen samples (n = 82) were examined to investigate whether the degree of sperm tail swelling in hypo-osmotic medium (fructose and sodium citrate; 150 mOsm/1), and motility characteristics after a 15-min exposure to hypotonic saline (sodium chloride; 150 mOsm/1) could predict the cryosurvival rate (% post-thaw motility/% pre-freeze motility x 100%) of spermatozoa after cryopreservation by the liquid nitrogen vapour freezing method using the TEST—glycerol—egg yolk buffer. The CellSoft automated semen analyser was used to analyse sperm motility in pre-freeze and post-thaw semen samples, and after exposure to hypotonic saline. Sperm tail hypo-osmotic swelling and sperm motility in pre-freeze semen showed no significant correlations (P > 0.05) with the cryosurvival rate. There were significant correlations (P < 0.05) between the cryosurvival rate and the following sperm motility parameters in hypotonic saline: % motility (r = 0.2846), motility index (% motility x curvilinear velocity; r = 0.2809) and % decrease in motility index from the baseline value in semen (r = 0.3378). The % decrease in motility index after hypotonic saline treatment was significantly less (P < 0.05) in the normal (≥50% cryosurvival rate; mean ± SEM 5.9 ± 3.2%; n = 33) compared with the subnormal (≥ 50% cryosurvival rate; 27.3 ± 4.8%; n = 49) cryosurvival groups. This parameter was also determined, by multivariate discriminant analysis, to be capable of classifying each prefreeze semen sample into either cryosurvival group with 69.5% accuracy. These results indicate that cryosurvival of spermatozoa is not dependent on pre-freeze motility characteristics, and that results of the tail hypo-osmotic swelling test cannot be utilized to predict the susceptibility of human spermatozoa to cryoinjury. Studies on the motility characteristics in an artificial medium such as hypotonic saline, while unable to provide a powerful predictive assay, may help to provide useful information about the survival of human spermatozoa after cryopreservation.

Key words: cryopreservation/human spermatozoa/hypo-osmotic swelling/hypotonic saline/motility


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