A simplified method for the isolation of sperm heads from the caput epididymidis of rodents and from ejaculated rabbit seminal plasma

Abstract

The present report describes a simple and rapid procedure to isolate highly purified sperm heads from mouse, rat and rabbit. The isolation procedure involves mincing of the caput epididymidis (CPE) of mice and rats to obtain a crude CPE suspension. This suspension is filtered to remove tubular fragments. The filtrate containing sperms and cellular contaminants is trypsinized, followed by sonication. To obtain rabbit sperm head suspension, ejaculated spermatozoa are briefly exposed to the cationic detergent CTAB-D, followed by trypsinization. Trypsin treatment of the suspension yields sperm heads and tails. The suspension containing sperm heads and tails is separated by discontinuous sucrose density gradient centrifugation. Highly purified and enriched sperm heads are recovered from the bottom of 2.5 M sucrose of the gradient. Enriched sperm heads contain less than 1% sperm tails.