Jiang Jin-Yi, Tsang Benjamin K
Reproductive Biology Unit and Division of Reproductive Medicine, Departments of Obstetrics & Gynecology and Cellular & Molecular Medicine, University of Ottawa, Ontario, Canada K1Y 4E9.
Biol Reprod. 2004 Dec;71(6):1974-9. doi: 10.1095/biolreprod.104.032839. Epub 2004 Aug 18.
The present study was conducted to determine the optimal conditions for successful in vitro fertilization (IVF) in Sprague-Dawley (SD) rats. The IVF of oocytes from SD and Wistar rats was compared in different fertilization media (mR1ECM, IVF-20, and modified Krebs-Ringer bicarbonate solution [mKRB]), and IVF conditions were then optimized for oocytes of the SD strain. Results showed that in mR1ECM medium, fertilization rates were markedly lower in SD rats (15%) than in the Wistar strain (73%), although this response was significantly improved by increasing the NaCl concentration. In addition, fertilization rates in SD rats were higher in modified IVF-20 (73%) than in IVF-20 (18%) and mKRB (53%). In contrast, fertilization rates in Wistar rats were higher in IVF-20 and modified IVF-20 than in mKRB (78%, 74%, and 36%, respectively). Further investigation concerning the effects of the NaCl supplementation (10- 40 mM) in IVF-20 on the fertilization of oocytes in the SD strain indicated that significantly higher percentages of oocytes were fertilized in IVF-20 supplemented with 30 mM NaCl (66%) and developed to the blastocyst stage (47%) in vitro. After transfer, embryos derived from this IVF system developed to term at a percentage comparable to that of in vivo-fertilized controls. In conclusion, differences exist in optimal IVF conditions between rat strains, and a modified culture medium has been successfully developed for assessment of the developmental competence of oocytes in SD rats.
本研究旨在确定斯普拉格-道利（SD）大鼠体外受精（IVF）成功的最佳条件。比较了在不同受精培养基（mR1ECM、IVF-20和改良的 Krebs-Ringer 碳酸氢盐溶液[mKRB]）中SD大鼠和Wistar大鼠卵母细胞的IVF情况，然后针对SD品系的卵母细胞优化IVF条件。结果显示，在mR1ECM培养基中，SD大鼠的受精率（15%）明显低于Wistar品系（73%），不过通过提高NaCl浓度，这一反应得到了显著改善。此外，SD大鼠在改良的IVF-20培养基中的受精率（73%）高于在IVF-20（18%）和mKRB（53%）中的受精率。相比之下，Wistar大鼠在IVF-20和改良IVF-20中的受精率高于在mKRB中的受精率（分别为78%、74%和36%）。关于在IVF-20中添加NaCl（10 - 40 mM）对SD品系卵母细胞受精影响的进一步研究表明，在添加30 mM NaCl的IVF-20中，受精的卵母细胞百分比显著更高（66%），并且在体外发育到囊胚阶段的百分比为47%。移植后，源自该IVF系统的胚胎发育至足月的百分比与体内受精对照组相当。总之，大鼠品系之间在最佳IVF条件方面存在差异，并且已成功开发出一种改良培养基用于评估SD大鼠卵母细胞的发育能力。
