Byers Shannon L, Wiles Michael V, Taft Robert A
Technology Evaluation and Development, The Jackson Laboratory, Bar Harbor, Maine, USA.
J Am Assoc Lab Anim Sci. 2009 Jan;48(1):44-51.
A novel surgical method for collecting oocytes from unique and irreplaceable mice is described. This method, surgical oocyte retrieval (SOR), facilitates the collection of ovulated oocytes, does not require euthanasia, and preserves reproductive potential. The surgery involves a small incision in the ampulla region of the oviduct, through which the cumulus oocyte mass is removed with a gel-loading pipette. The incision then is closed by using a tissue adhesive, which is required to ensure healing of the incision and containment of any oocytes ovulated after SOR. Two anesthetics, isoflurane and tribromoethanol, were compared for oocyte toxicity during SOR. More dead oocytes were recovered when tribromoethanol was used than when isoflurane was used. Combining SOR and traditional oocyte collection methods yielded more oocytes per BALB/cByJ than did traditional methods alone (41 versus 28 oocytes, respectively). Oocytes collected by using SOR were fertilized and subsequent embryos developed to term comparable to controls. This technique provides an alternative method for oocyte collection and will be valuable for maximizing the number of oocytes from irreplaceable mice.
本文描述了一种从独特且不可替代的小鼠体内采集卵母细胞的新型手术方法。这种方法,即手术取卵(SOR),便于收集排卵后的卵母细胞,无需实施安乐死,并保留了生殖潜力。手术包括在输卵管壶腹部区域做一个小切口,通过这个切口用凝胶上样移液器取出卵丘卵母细胞团。然后用组织粘合剂封闭切口,这对于确保切口愈合以及防止SOR后排出的任何卵母细胞外溢是必需的。比较了两种麻醉剂异氟烷和三溴乙醇在SOR过程中对卵母细胞的毒性。使用三溴乙醇时回收的死亡卵母细胞比使用异氟烷时更多。与单独使用传统方法相比,将SOR与传统卵母细胞采集方法相结合,每只BALB/cByJ小鼠获得的卵母细胞更多(分别为41个和28个卵母细胞)。通过SOR采集的卵母细胞受精后,随后的胚胎发育至足月,与对照组相当。这项技术为卵母细胞采集提供了一种替代方法,对于最大限度地从不可替代的小鼠体内获取卵母细胞具有重要价值。