Esteves S C, Sharma R K, Thomas A J, Agarwal A
Department of Urology, Cleveland Clinic Foundation, Ohio 44195, USA.
Fertil Steril. 1996 Nov;66(5):798-804.
To determine whether the hypo-osmotic swelling test can assess viability of frozen spermatozoa.
Fresh semen specimens were divided into two aliquots: one was not processed and the other was processed by swim-up. Both aliquots were then cryopreserved by the liquid nitrogen vapor method.
Andrology laboratory at a tertiary care facility.
PATIENT(S): Eleven volunteers with proven fertility.
INTERVENTION(S): The hypo-osmotic swelling test and the sperm viability assessment by Hoechst 33258 (Sigma Chemical Co., St. Louis, MO) and eosin-nigrosin strains were performed in each aliquot.
MAIN OUTCOME MEASURE(S): The results of hypo-osmotic swelling test as indicated by swelling of sperm tails were compared with the viability results obtained by Hoechst 33258 stain before freezing and by Hoechst-33258 and eosin-nigrosin stains after freezing.
RESULT(S): The hypo-osmotic swelling test and Hoechst 33258 viability results were correlated highly in unprocessed and processed fresh specimens (r = 0.95). After cryopreservation, there was no correlation between hypo-osmotic swelling test and Hoechst 33258 or between hypo-osmotic swelling test and eosin-nigrosin. Prefreeze sperm processing did not influence the post-thaw results. Viability assessed by Hoechst 33258 and eosin-nigrosin was well correlated (r = 0.72).
CONCLUSION(S): The hypo-osmotic swelling test can accurately evaluate viability in fresh human spermatozoa but not in cryopreserved ones. Thus, this test cannot be used to select viable cells in cryopreserved samples for assisted reproductive procedures, such as intracytoplasmic sperm injection.
确定低渗肿胀试验能否评估冷冻精子的活力。
将新鲜精液标本分成两份：一份不做处理，另一份采用上游法处理。然后两份标本均采用液氮蒸汽法冷冻保存。
一家三级医疗机构的男科实验室。
11名经证实有生育能力的志愿者。
对每份标本进行低渗肿胀试验，并用Hoechst 33258（西格玛化学公司，密苏里州圣路易斯）、伊红-黑色素染色法评估精子活力。
将精子尾部肿胀所显示的低渗肿胀试验结果，与冷冻前用Hoechst 33258染色及冷冻后用Hoechst 33258和伊红-黑色素染色所获得的活力结果进行比较。
在未经处理和经处理的新鲜标本中，低渗肿胀试验结果与Hoechst 33258活力结果高度相关（r = 0.95）。冷冻保存后，低渗肿胀试验结果与Hoechst 33258结果之间、低渗肿胀试验结果与伊红-黑色素结果之间均无相关性。冷冻前的精子处理不影响解冻后的结果。用Hoechst 33258和伊红-黑色素评估的活力相关性良好（r = 0.72）。
低渗肿胀试验可准确评估新鲜人类精子的活力，但不能评估冷冻精子的活力。因此，该试验不能用于在冷冻保存的样本中选择有活力的细胞用于辅助生殖程序，如卵胞浆内单精子注射。
