摘要
In vitro fertilisation (IVF) protocols for red deer have yielded low fertilisation rates, with no embryo development beyond the eight-cell stage when heparin was used as the in vitro capacitation agent. As this low fertilisation rate may result from reduced motility, the present study investigated the use of red deer oviduct epithelial cell monolayers (COEM) and conditioned medium (Cm) from the monolayers to maintain red deer sperm motility in vitro. A second experiment compared the fertilisability of red deer sperm pre-incubated for 4–12 h on COEM or for 4 h in TALP medium supplemented with 20 μg of heparin.
COEM was superior in maintaining red deer sperm motility compared with either Sp-TALP alone or Cm (P h period. The motility of sperm incubated in Sp-TALP and Cm was similar and had declined to less than 10% by 4 h and no motile sperm were observed by 8 h. Overall, the penetration rates of in vitro red deer oocytes were low (5–28%) regardless of sperm treatment. Sperm pre-incubated on COEM penetrated more oocytes than sperm incubated with heparin (P h pre-incubation of sperm on COEM (P0.50). Penetration rates were greater across all treatments when both sperm and oocytes were co-incubated for 24 h compared to 12 h (P
It was concluded that COEM sustains red deer sperm motility in vitro during the 24 h observation period. Pre-incubating sperm on COEM does increase sperm penetration rates compared with heparin alone, but at a rate too low and variable to be used on a routine basis. Overall, the penetration rates were comparable to those previously reported for red deer even though differences in heparin concentration, fertilisation systems and stags were used.
摘要译文
马鹿体外受精（IVF）方案的受精率较低，当使用肝素作为体外获能剂时，超过八细胞阶段没有胚胎发育。由于这种低受精率可能是由于运动力减弱所致，本研究调查使用来自单层的马鹿输卵管上皮细胞单层（COEM）和条件培养基（Cm）维持体外的马鹿精子活力。第二个实验比较了在鹿精子上预孵育4-12小时的马鹿精子的受精能力，或在补充有20μg肝素的TALP培养基中4小时的受精能力。与单独的Sp-TALP或Cm（P期）相比，COEM在维持马鹿精子活力方面是优越的。Sp-TALP和Cm孵育的精子运动能力相似，在4小时内下降到10％以下，8小时没有运动精子。总体，无论精子处理如何，体外红鹿卵母细胞的穿透率都很低（5-28％）。在COEM上预孵育的精子比用肝素孵育的精子渗透更多的卵母细胞（在COEM上预孵育精子（P0.50））。当精子和卵母细胞共同孵育24小时时，与12小时相比，所有处理都是一样的（P结论是COEM在24小时观察期间维持了体外马鹿精子活力。在COEM上预孵育精子确实可以提高精子穿透率，与单独使用肝素相比，但是速度太低，并且可以根据常规使用。总体，即使使用肝素浓度，施肥系统和雄鹿的差异，穿透率也与以前报道的马鹿的穿透率相当。
D.KBerg[a][c];J.GThompson[a][1];G.WAsher[b];. Development of in vitro embryo production systems for red deer (Cervus elaphus)[J]. Animal Reproduction Science, 2002,70(1-2): 65-76