人睾丸组织中成熟精子的分离方法初探  

Primary Study on the Method for Separating Mature Sperms from Human Testicular Tissue

作　　者：李培旭[1] 李尚为[1] 彭芝兰[1] 马黔红[1] 黄仲英[1] 李蕾[1] 唐宗蓉[1] 韩字研[1] 

机构地区：[1]四川大学华西第二医院妇产科,成都610041

出　　处：《生物医学工程学杂志》2005年第2期371-373,共3页Journal of Biomedical Engineering

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。This study was aimed to explore a simple and applicable method of separating mature sperms from human testicular tissue for intracytoplasmic sperm injection and embryo transfer. The suspension of human testicular tissue 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 centrifugation was processed to separate the sperms ; meanwhile the sperms were counted and graded according to their motility. The difference in quality and quantity among the groups and the difference between the groups and the zero-hour culturing group were detected. It was shown that the four hyaluronidase-treated groups contained large quantity and high quality of sperms as compared with the two contrast groups（P〈0.01）. The groups in the solution of 50u/ml, 100u/ml and 150u/ml concentrations of hyaluronidase had almost the same amount of sperms that displayed higher motility as compared against the sperms in the group treated with 200u/ml concentration of hyaluronidase （P〈0. 01）. There was no difference between the two contrast groups （P〈0.05）, or among the groups treated with 50u/ml, 100u/ml, and 150u/ml of hyaluronidase concentration （P〉0.05）. This method of adopting hyaluronidase with Percoll gradient centrifugation in the process for separating mature sperms from human testicular tissue is applicable. It can increase the quantity and quality of sperms separated from testicular tissue suspensions when adequate concentrations of hyaluronidase is used.

关 键 词：睾丸精子 透明质酸酶 Percoll梯度离心 成熟精子 睾丸组织 Percoll梯度离心法 分离方法 卵母细胞内单精子注射 睾丸曲细精管 人血白蛋白 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学]