Harrison R A, Dott H M, Foster G C
J Exp Zool. 1982 Jul 20;222(1):81-8. doi: 10.1002/jez.1402220111.
Epididymal spermatozoa from rabbit and ram were washed either once or twice using an efficient washing procedure and were then diluted in various media to a final concentration of approximately 1.4 x 10(7) cells/ml and incubated at 30 degrees C for up to 12 hours. Bovine serum albumin (BSA), either untreated or defatted, was found to be better than polyvinylpyrrolidone, ovalbumin, or alpha-lactalbumin, both at stimulating and maintaining motility levels and at reducing the tendency of the washed spermatozoa to stick to glass. BSA was effective in all media tested, being independent of Ca2+, PO4(3-), HCO3-, and ionic strength). BSA has a reversible stimulatory effect on motility. If BSA was added to sperm suspensions 3 1/2 hours after they had been washed and diluted in protein-free medium, motility was stimulated to levels not significantly lower than those observed in samples that had been washed and diluted in the presence of BSA. However, samples washed into BSA and then washed free of it behaved essentially as though they had never been in contact with protein. The motility, survival, and response to BSA of twice-washed spermatozoa were the same as those of once-washed spermatozoa, showing that epididymal plasma factors are not required for survival in vitro. It was concluded that dilution is not essentially detrimental to rabbit and ram spermatozoa. However, severe dilution of semen may result in levels of male reproductive tract fluids insufficient either to stimulate motility or to prevent sticking of motile cells to container surfaces. Few motile spermatozoa are recovered from samples of such diluted semen.
来自兔和公羊的附睾精子，使用高效洗涤程序洗涤一次或两次，然后在各种培养基中稀释至最终浓度约为1.4×10⁷个细胞/毫升，并在30℃下孵育长达12小时。发现未处理或脱脂的牛血清白蛋白（BSA）在刺激和维持活力水平以及减少洗涤后的精子粘附于玻璃的趋势方面，均优于聚乙烯吡咯烷酮、卵清蛋白或α-乳白蛋白。BSA在所有测试的培养基中均有效，且与Ca²⁺、PO₄³⁻、HCO₃⁻和离子强度无关。BSA对活力有可逆的刺激作用。如果在精子悬浮液在无蛋白培养基中洗涤和稀释3.5小时后加入BSA，活力会被刺激到不显著低于在有BSA存在下洗涤和稀释的样品中观察到的水平。然而，先洗涤到BSA中然后再洗涤去除BSA的样品，其表现基本上就好像它们从未接触过蛋白质一样。两次洗涤的精子的活力、存活率和对BSA的反应与一次洗涤的精子相同，表明体外存活不需要附睾血浆因子。得出的结论是，稀释对兔和公羊精子基本上无害。然而，精液的严重稀释可能导致雄性生殖道液水平不足以刺激活力或防止活动细胞粘附于容器表面。从这种稀释精液的样品中回收的活动精子很少。
