Predictive value of chromatin decondensation in vitro on fertilization rate after intracytoplasmic sperm injection (ICSI).
This study was undertaken to identify the relationship between sperm chromatin decondensation after incubation with sodium dodecyl sulphate (SDS) and ethylene diamine tetra acetic acid (EDTA), or heparin at various points of time. Likewise, this study will determine chromatin stability within definite time intervals, chromatin decondensation after intracytoplasmic sperm injection (ICSI), and whether chromatin decondensation in vitro could be used as a predictive test for fertilization capability after ICSI. Sixty-five infertile couples undergoing ICSI therapy were included in this prospective study. Male factor infertility was the main indication for inclusion. One millilitre from each semen sample after washing was mixed with SDS-EDTA (group 1) or SDS/heparin (group 2) and incubated for 120 min. Many smears were made within 10 min of mixing the spermatozoa with detergent and the reducing agents and at the following points of time 30, 60 and 120 min and after 24 h. Chromatin decondensation was evaluated after staining with acridine orange (AO). The mean percentage of uncondensed chromatin of spermatozoa in the semen sample in the first group before addition of SDS/EDTA was 26.1 +/- 19.0 and 22.3 +/- 18.9% in the second one. After incubation of spermatozoa for 30, 60 and 120 min and 24 h, the chromatin decondensation increased in the first group to 64.0 +/- 28.6, 83.0 +/- 21.1, 87.9 +/- 14.6, 92.1 +/- 16.2 and 98.0 +/- 6.75%, respectively. The corresponding values in the second group were 69.5 +/- 29.9, 78.6 +/- 22.4, 86.9 +/- 17.1, 95.13 +/- 6.5 and 98.3 +/- 5.6%. On the other hand, no correlations were found between the chromatin decondensation or chromatin decondensation rate in vitro and the fertilization rates in all investigated groups. In conclusion, neither the chromatin decondensation ability in vitro nor the rate of chromatin decondensation between various points of time after using SDS/EDTA, SDS/heparin could predict the chromatin decondensation of spermatozoa (fertilization capability) after ICSI.
Hammadeh
M E
ME
Department of Obstetrics and Gynaecology, Medical College, University of Saarland, 66421 Homburg/Saar, Germany. frmham@med-rz.uni-sb.de
Al-Hasani
S
S
Gauss
C
C
Rosenbaum
P
P
Georg
T
T
Diedrich
K
K
Schmidt
W
W
D000328:Adult; D002843:Chromatin / Q000502:physiology*; D004492:Edetic Acid / Q000494:pharmacology; D005260:Female; D005298:Fertility; D005306:Fertilization; D006493:Heparin / Q000494:pharmacology; D006801:Humans; D008297:Male; D011237:Predictive Value of Tests; D011247:Pregnancy; D011446:Prospective Studies; D012967:Sodium Dodecyl Sulfate / Q000494:pharmacology; D020554:Sperm Injections, Intracytoplasmic; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology*
D002843:Chromatin; D012967:Sodium Dodecyl Sulfate; D006493:Heparin; D004492:Edetic Acid