Establishment of an optimal hypo-osmotic swelling test by examining single spermatozoa in four different hypo-osmotic solutions.
In order to find an optimal hypo-osmotic swelling test (HOST) and to identify viable sperm cells from patients with asthenozoospermia for intracytoplasmic sperm injection (ICSI), we tested single motile and non-motile spermatozoa in four hypo-osmotic solutions by micromanipulation. The four solutions were: A, H2O; B, 50 mOsm NaCl; C, 150 mOsm NaCl and D, 150 mOsm sodium citrate and fructose. Eosin Y staining was then carried out for evaluation of viability of the spermatozoa after HOST. Using motile spermatozoa, no statistical difference was found in HOST-positive spermatozoa between these four solutions. There were more viable sperm cells after HOST in solutions C and D, as noted by Eosin Y staining. After non-motile spermatozoa were incubated for 1 min in the four solutions, HOST with solution C gave the best results for identification of viable sperm cells compared to the other three solutions. When motile spermatozoa were incubated in solution C or solution D for 30 min, the result of HOST with solution C (10.8% dead spermatozoa) was superior to that of solution D (49.1% dead spermatozoa). In conclusion, the HOST protocol using 150 mOsm NaCl (solution C) for 1 min yielded the best results for selection of viable spermatozoa. This procedure should be used for selection of viable spermatozoa for ICSI in patients with 100% non-motile spermatozoa.
Tsai
Y L
YL
The Department of Obstetrics and Gynecology, Shin Kong Wu Ho-Su Memorial Hospital, Shin Lin, Taipei, Taiwan.
Liu
J
J
Garcia
J E
JE
Katz
E
E
Compton
G
G
Baramki
T A
TA
D002462:Cell Membrane / Q000502:physiology; D048429:Cell Size; D002470:Cell Survival; D004801:Eosine Yellowish-(YS); D005456:Fluorescent Dyes; D006801:Humans; D007038:Hypotonic Solutions / Q000737:chemistry; D008297:Male; D009994:Osmolar Concentration; D012661:Semen / Q000166:cytology / Q000502:physiology; D012680:Sensitivity and Specificity; D013081:Sperm Motility / Q000502:physiology*; D013094:Spermatozoa / Q000166:cytology / Q000502:physiology*
D005456:Fluorescent Dyes; D007038:Hypotonic Solutions; D004801:Eosine Yellowish-(YS)