摘要
目的探讨分离、纯化人睾丸精原细胞的有效方法。方法用联合二步酶消化法获得人生殖细胞悬液,经Percoll不连续密度梯度离心,再用间隔不同时间(2、3、4h),不同次数(1、2次)差异黏附法纯化人精原细胞。结果未经纯化的精原细胞纯度为48.5%,间隔2、3、4h纯化1次所获得的精原细胞纯度分别为51.3%、59.0%、56.4%,纯度均高于未经纯化的纯度;间隔2、3、4h连续纯化2次所获精原细胞纯度分别为:57.7%、67.8%、57.8%,以间隔3h纯度最高;与间隔同样时间纯化1次精原细胞纯度相比,间隔2、3h纯化2次高于1次,间隔4h纯化2次与1次无明显差别。结论Percoll不连续密度梯度离心联合不同时间间隔差异黏附法可以有效地对人精原细胞进行分离和纯化;间隔2、3h,纯化2次高于1次;连续纯化2次时,间隔3h效果最佳。
Objective To research the effective methods of separation and purification of human testicle spermatogonia. Methods The two-step enzymatic digestion was used to obtain germ cells suspension to be centrifuged by Perco11 discontinuous density gradient centrifugation. Finally, purified human spermatogonias were obtained according to different time and times of cellular adhesiveness （per 2, 3, 4 h and one or twp times）. Results （1）The percentage of spermatogonia only after separated by Percoll was 48.5% ,and that of purified one time by interval 2, 3, 4 h was S 1.3%, 59.0%, 56.4% respectively; （2）The percentage of spermatogonia was 57.7%, 67.8%, 57.8% respectively through 2, 3, 4 h interval and continuous 2 times; The highest was in interval of 3 h;（3）Compared with purified one time,that in interval of 2, 3 h and two times were higher than that of one time; But that of 4 h and two times had no difference from that of one time. Conclusions （1）Percoll centrifugation combined with different time of cellular adhesiveness can effectively separate and purify the human spermatogonia.（2）The percentage of spermatogonia of purification by two times is higher than that of one time by interval of 2, 3 h; （3）If purification by continuous two times, interval 3 h is most effective.
出处
《中国老年学杂志》
CAS
CSCD
北大核心
2007年第2期117-119,共3页
Chinese Journal of Gerontology
基金
吉林省科技厅科研基金资助(20050401-1)
关键词
人
精原细胞
分离
纯化
Human spermatogonia
Separation
Purification
