Bongso A, Trounson A
Human and Animal Reproductive Biology Laboratories, Institute of Reproduction and Development, Monash University, Clayton, Victoria, Australia.
Hum Reprod. 1996 Jul;11(7):1451-6. doi: 10.1093/oxfordjournals.humrep.a019418.
Murine sperm from the caput, corpus and cauda epididymis were cocultured with epididymal epithelial cells of their own region or more distal regions, in the presence and absence of androgens (testosterone and dihydrotestosterone). Epithelial cell cultures were used 3 or 10 days after preparation in a complex tissue culture medium (Chang's) as plated tubules. The coculture studies involving spermatozoa and oocytes with epithelial cells were carried out in T6 medium. Motility of caput spermatozoa was maintained for 24 h in the presence of day 3 corpus and cauda epithelial cells and hormones but not under other conditions. Likewise, the motility of corpus spermatozoa was maintained for 24 h in the presence of day 3 cauda epithelial cells and hormones but not other conditions. Fertilization of zona-intact oocytes by epididymal spermatozoa was not affected by their coculture for 24 h with epithelial cells but fertilization rates for zone-free oocytes were increased for caput spermatozoa cocultured with more distal epithelial cells. Fertilization rates for both zona-intact and zone-free oocytes were increased for corpus spermatozoa cocultured with more distal cauda epithelial cells. The developmental capacity of embryos derived from caput spermatozoa was not significantly increased by coculture with epithelial cells but those derived from corpus spermatozoa cocultured with cauda epithelial cells were significantly increased. We conclude that the presence of more distal epithelial cells of the mouse epididymis maintains motility in culture, increases the ability of caput and corpus spermatozoa to fertilize zona-free oocytes and increases the developmental capacity of embryos formed from corpus spermatozoa. These observations demonstrate the function of epididymal regions in the maturation of murine spermatozoa for fertilization and embryo development.
将来自附睾头、体和尾的小鼠精子与自身区域或更远端区域的附睾上皮细胞进行共培养,培养过程中添加或不添加雄激素(睾酮和双氢睾酮)。上皮细胞培养物在制备后3天或10天,以铺板小管的形式用于复杂组织培养基(张氏培养基)中。涉及精子和卵母细胞与上皮细胞的共培养研究在T6培养基中进行。附睾头精子在第3天的附睾体和尾上皮细胞及激素存在的情况下,其活力可维持24小时,但在其他条件下则不能。同样,附睾体精子在第3天的附睾尾上皮细胞及激素存在的情况下,其活力可维持24小时,但在其他条件下则不能。附睾精子对完整透明带卵母细胞的受精不受与上皮细胞共培养24小时的影响,但对于与更远端上皮细胞共培养的附睾头精子,去透明带卵母细胞的受精率有所提高。对于与更远端附睾尾上皮细胞共培养的附睾体精子,完整透明带和去透明带卵母细胞的受精率均有所提高。附睾头精子来源的胚胎的发育能力通过与上皮细胞共培养没有显著提高,但附睾体精子与附睾尾上皮细胞共培养来源的胚胎的发育能力显著提高。我们得出结论,小鼠附睾更远端上皮细胞的存在可维持培养中的精子活力,提高附睾头和附睾体精子对去透明带卵母细胞的受精能力,并提高由附睾体精子形成的胚胎的发育能力。这些观察结果证明了附睾区域在小鼠精子成熟以进行受精和胚胎发育中的作用。