Ohashi K, Saji F, Wakimoto A, Tsutsui T, Nakazawa T, Okabe M, Mimura T, Tanizawa O
Department of Obstetrics and Gynecology, Osaka University Medical School, Japan.
J Androl. 1994 Jan-Feb;15(1):78-82.
Separation of acrosome-reacted viable sperm is an important problem in the analysis of sperm function and the micromanipulation of gametes such as the microinjection of sperm into ooplasm and subzonal sperm insertion. In the present study, we collected acrosome-reacted sperm selectively using paramagnetic immunobeads, referred to as MH61-beads, which attach themselves to the head of the acrosome-reacted sperm and are recovered by a magnet. The maximum number of sperm was recovered by the MH61-beads when 3 x 10(5) sperm/100 microliters and 6 x 10(5) MH61-beads were coincubated for 1 hour. The recovery of sperm was significantly correlated with the percentage of the acrosome reaction in the capacitation medium. After incubation for 12 hours, 2.4-9.0 x 10(4) sperm could be collected, and the percentage of viable and acrosome-reacted sperm was 28-60%. To improve the recovery of the acrosome-reacted sperm, we utilized follicular fluid, which induces the acrosome reaction. Addition of follicular fluid to the capacitation medium at a final concentration of 20% induced the acrosome reaction and increased the recovery of acrosome-reacted sperm significantly. In conclusion, separation of acrosome-reacted sperm by the MH61-bead-binding method could become a useful technique in research of sperm function and gamete micromanipulation.
顶体反应后活精子的分离是精子功能分析以及配子显微操作（如将精子显微注射到卵质和透明带下精子注入）中的一个重要问题。在本研究中，我们使用顺磁性免疫磁珠（称为MH61磁珠）选择性收集顶体反应后的精子，这些磁珠会附着在顶体反应后精子的头部，并通过磁铁回收。当3×10⁵个精子/100微升与6×10⁵个MH61磁珠共同孵育1小时时，通过MH61磁珠回收的精子数量最多。精子的回收率与获能培养基中顶体反应的百分比显著相关。孵育12小时后，可收集到2.4 - 9.0×10⁴个精子，活的且发生顶体反应的精子百分比为28% - 60%。为了提高顶体反应后精子的回收率，我们利用了诱导顶体反应的卵泡液。在获能培养基中添加终浓度为20%的卵泡液可诱导顶体反应，并显著提高顶体反应后精子的回收率。总之，通过MH61磁珠结合法分离顶体反应后的精子可能成为精子功能研究和配子显微操作中的一项有用技术。
