FERTILIZING ABILITY OF BOVINE SPERMATOZOA COCULTURED WITH OVIDUCT EPITHELIAL-CELLS
FERTILIZING ABILITY OF BOVINE SPERMATOZOA COCULTURED WITH OVIDUCT EPITHELIAL-CELLS
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DOI：
10.1095/biolreprod52.1.156
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发表时间：
1995-01-01
影响因子：
3.6
通讯作者：
SIRARD, MA
中科院分区：
文献类型：
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作者：
CHIAN, RC;SIRARD, MA
采用体外细胞培养技术，研究了不同条件下不同部位的牛输卵管上皮细胞单层（OECM）对牛精子穿透卵母细胞的影响。用OECM+ OECM条件培养基处理的精子的卵母细胞穿透率为93%（155/166）。OECM+新鲜培养基和单独OECM条件培养基中的精子穿透率分别为66%（115/173）和52%（87/167），显著低于（p < 0.01）OECM+ OECM条件培养基。然而，在单独的OECM条件培养基中穿透卵母细胞的百分比显著高于对照组（11% = 16/148;p < 0.01）。在OECM+ OECM条件培养液组和OECM+新鲜培养液组中，与来自峡部段的OECM共培养的精子的卵母细胞穿透率显著低于与来自壶腹段的OECM共培养的精子的卵母细胞穿透率（分别为44% = 43/98 vs. 72% = 68/94，42% = 49/117 vs. 66% = 72/110; p < 0.01）。精子穿透率低，在OECM条件培养基单独来自峡部或壶腹段（20% = 20/100和19% = 17/81，分别）。而从全输卵管OECM中获得的OECM条件培养液可显著提高精子穿透率（57% = 60/105;p <0.01）。体外研究了不同节段OECM对精子结合和维持精子运动能力的影响。共培养2小时后，超过一半的精子附着在OECM上，无论其来源如何。共培养3 h后，整个输卵管和壶腹段的OECM中逐渐释放精子;然而，共培养12 h后，精子仍然附着在峡部OECM上。共培养48 h后，峡部OECM或全输卵管OECM中未附着精子的活动率显著高于壶腹OECM（分别为52% + 3、55% +/- 4和25% +/- 4; P < 0.01）。当精子与壶腹OECM共培养时，从共培养后24小时起，活动精子的百分比显著低于与全输卵管OECM和峡部OECM共培养后获得的百分比（分别为26% +/- 4、63% +/- 3和61% +/- 4; p < 0.01）。这些结果表明，精子获能是协同诱导的附着到OECM和暴露于OECM条件培养基的装置，和精子附着到壶腹上皮增强精子获能。这些结果还表明，峡部上皮细胞是重要的，在维持精子的活力和附着的精子在峡部可能会减少精子到达壶腹在体内的数量。
The effects of bovine oviduct epithelial cell monolayers (OECM), derived from the different segments of the oviduct and under various conditions, on oocyte penetration by bovine sperm were determined by in vitro cell culture techniques. The oocyte penetration rate by sperm treated with OECM+OECM-conditioned medium was 93% (155 of 166). Sperm penetration rates in OECM+fresh medium and in OECM-conditioned medium alone were 66% (115 of 173) and 52% (87 of 167), respectively, significantly lower (p < 0.01) than in OECM+OECM-conditioned medium. However, the percentage of penetrated oocytes in OECM-conditioned medium alone was significantly higher than in control (11% = 16 of 148;p < 0.01). In both the OECM+OECM-conditioned medium and the OECM+fresh medium groups, oocyte penetration rates by sperm cocultured with OECM derived from the isthmic segment were significantly lower than those by sperm cocultured with OECM derived from the ampullary segment (44% = 43 of 98 vs. 72% = 68 of 94, and 42% = 49 of 117 vs. 66% = 72 of 110, respectively; p < 0.01). Sperm penetration rates were low after insemination in the OECM-conditioned medium alone derived from either the isthmic or the ampullary segments (20% = 20 of 100 and 19% = 17 of 81, respectively). However, the sperm penetration rate was improved significantly when OECM-conditioned medium was obtained from whole-oviduct OECM (57% = 60 of 105;p < 0 01). The effect of OECM derived from different segments on ability of sperm binding and maintaining motility was also evaluated in vitro. After 2 h of coculture, more than half the sperm attached to OECM regardless of their origin. Sperm were gradually released from OECM in the whole oviduct and ampullary segments after 3 h of coculture; however, sperm remained attached to isthmic OECM after 12 h of coculture. After 48 h of coculture, the motility of unattached sperm in either isthmic OECM or whole-oviduct OECM was significantly higher than in ampullary OECM (52% + 3, 55% +/- 4, and 25% +/- 4, respectively; P < 0.01). When sperm were cocultured with ampullary OECM, the percentage of motile sperm was significantly lower from 24 h after coculture than percentages obtained after coculture with whole-oviduct OECM and isthmic OECM (26% +/- 4, 63% +/- 3, and 61% +/- 4, respectively; p < 0.01). These results suggest that sperm capacitation was synergistically induced by means of both attachment to OECM and exposure to OECM-conditioned medium, and that sperm attachment to the ampullary epithelium enhanced sperm capacitation. These results also suggest that the isthmus epithelial cells are important in maintaining spertn motility and that attachment of sperm in the isthmus may act to reduce the number of sperm arriving at the ampulla in vivo.