【摘要】 探索简单、适用的体外分离睾丸组织中成熟精子的方法,以用于男性无精子症病人行卵母细胞内单精子注射受精-胚胎移植为目的。用含不同浓度透明质酸酶的10 %人血白蛋白人类输卵管液培养人睾丸曲细精管组织匀浆2 4 h,再用Percoll梯度离心法洗涤后计算所获各级精子的总数,并设置前后对照和空白对照进行比较。用不同浓度透明质酸酶处理后的4个实验组分别与2个对照组比较,实验组所获得的各级精子的总数高于对照组,有显著差异(P<0 .0 1)。实验组中透明质酸酶浓度分别为5 0、10 0、15 0 u/ ml的10 %人血白蛋白人类输卵管液处理组获得的各级精子总数组间比较,处理前对照和处理后空白对照之间比较,无显著差异(P>0 .0 5 )。而用浓度为2 0 0 u/ ml的10 %人血白蛋白人类输卵管液处理组获得的各级精子的总数与其它浓度处理组之间比较存在统计学差异(P<0 .0 1) ,其所获得的各级精子总数相似,但活动能力较其它浓度处理组低。透明质酸酶结合Percoll梯度离心可从人睾丸曲细精管组织匀浆中分离出较多更纯的成熟精子,但过高浓度的透明质酸酶可能会影响分离出的精子的活动能力。更多还原

【Abstract】 This study was aimed to explore a simple and applicable me thod of separating mature sperms from human testicular tissue for intracytoplasm ic sperm injection and embryo transfer. The suspension of human testicular tissu e was cultured in 10% human serum albumin and human tubule fluid with different concentrations(0u/ml; 50u/ml; 100u/ ml; 150u/ml; 200u/ml) of hyaluronidase for 24h, and then the Percoll gradient ce ntrifugation was processed to separate the sperms; meanwhile the sperms were cou n ted and graded according to their motility. The difference in quality and quanti ty among the groups and the difference between the groups and the zero-hour culturing group were detected. It was shown that the four hyaluronidase-treate d groups contained large quantity and high quality of sperms as compared with th e two contrast groups(P<0.01). The groups in the solution of 50u/ml, 100 u/ml and 150u/ml concentrations of hyaluronidase had almost the same amount of s perms that displayed higher motility as compared against the sperms in the grou p treated with 200u/ml concentration of hyaluronidase (P<0.01). There was n o difference between the two contrast groups(P>0.05), or among the groups tr eated with 50u/ml, 100u/ml, and 150u/ml of hyaluronidase concentration (P>0.05). This method of adopting hyaluronidase with Percoll gradient centr ifugation in the process for separating mature sperms from human testicula r tissue is applicable. It can increase the quantity and quality of sperms separ ated from testicular tissue suspensions when adequate concentrations of hyaluron idase is used.更多还原