上海交通大学学报（医学版）

›› 2012, Vol. 32 ›› Issue (8): 979-.doi: 10.3969/j.issn.1674-8115.2012.08.004

• 专题报道（不孕不育症及辅助生殖技术） • 上一篇    下一篇

温度和Hoechst 33258浓度对人活精子标记效果的影响

卢 慧, 施文博, 肖玉芳, 王柱清, 刘 勇, 卢克敏, 汪小波, 李 铮   

  1. 上海交通大学 医学院附属仁济医院泌尿外科 |上海市人类精子库 上海市男科学研究所精子发育与遗传实验室, 上海 200001
  • 出版日期:2012-08-28 发布日期:2012-08-29
  • 通讯作者: 李 铮, 电子信箱: Lizhengboshi@163.com。
  • 作者简介:卢 慧（1978—）, 女, 实习研究员, 硕士；电子信箱: lzaloe@sina.com。
  • 基金资助:

    上海市科委重点课题（10JC1409900）

Influence of temperature and Hoechst 33258 concentration on vital labeling of human sperm

LU Hui, SHI Wen-bo, XIAO Yu-fang, WANG Zhu-qing, LIU Yong, LU Ke-min, WANG Xiao-bo, LI Zheng   

  1. Shanghai Human Sperm Bank, Department of Urology, Sperm Development and Genetics Laboratory, Shanghai Institute of Andrology, Renji Hospital, Shanghai Jiaotong University School of Medicine, Shanghai 200001, China
  • Online:2012-08-28 Published:2012-08-29
  • Supported by:

    Shanghai Science and Technology Committee Foundation,10JC1409900

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摘要：

目的 研究孵育温度和荧光染料Hoechst 33258（H258）的浓度对活体标记人类精子效果的影响。方法 随机选取14份精液标本，经洗涤处理后，在不同温度（0 ℃、室温和37 ℃）下分别用0（对照）、0.01、0.1 mg/mL的H258孵育1 h。 记录精子前向运动百分率及H258着色率。以未予处理的精液标本作为空白对照组。结果 在观察条件范围内，孵育温度与H258浓度对精子运动的影响有统计学意义。随着H258质量浓度的升高和孵育温度的降低，精子前向运动百分率下降（P<0.01）。精子着色率随孵育温度和H258质量浓度的升高而提高；当H258质量浓度为0.1 mg/mL时，不同孵育温度下的精子着色率比较差异有统计学意义（P<0.01）。结论 H258可用作精子活体荧光标记，37 ℃+0.1 mg/mL H258是本研究中最优的染色条件。但仍需要根据不同使用目的选择孵育条件。

关键词: 精子, 温度, Hoechst 33258, 活体标记

Abstract:

Objective To investigate the influence of culture temperature and concentration of fluorescence dye Hoechst 33258 (H258) on vital labeling of human sperm. Methods Fourteen semen samples were randomly selected, and were cultured with 0 mg/mL (control), 0.01 mg/mL and 0.1 mg/mL H258 for 1 h under different temperatures (0 ℃, room temperature and 37 ℃) after washing. Progressive motility of human sperm was assessed, and H258 staining rates of the sperm were recorded. Semen samples without treatment were served as blank controls. Results Under the conditions of this research, H258 concentrations and culture temperatures had significant impact on sperm motility. The percents of progressive motility of sperm gradually decreased with the increase of H258 concentrations and decrease of culture temperatures (P<0.01). The staining rates of the sperm gradually increased with the culture temperatures and mass concentrations of H258, and there were significant differences among staining rates of the sperm cultured with different temperatures at the mass concentration of 0.1 mg/mL (P<0.01). Conclusion H258 can be used in vital labeling of human sperm. The optimal staining condition was 37 ℃+0.1 mg/mL H258 in this study. Culture condition should be determined by specific purpose.

Key words: sperm, temperature, Hoechst 33258, vital labeling