Similar patterns of embryo development in canine oocytes cultured in vitro at oxygen tensions of 5 and 20%

Abstract

The influence of two oxygen tensions (5 and 20%) on fertilization, cleavage, development, and morphological quality of canine embryos produced in vitro was investigated. To assess embryo production, canine oocytes (N = 608) were matured in vitro at 20% oxygen tension in TCM 199 supplemented with glucose (11 mM) and 0.1% polyvinyl alcohol. Oocytes and sperm were coincubated at 37 °C for 24 hours in serum-free medium. Subsequently, presumptive zygotes were cultured in vitro in synthetic oviductal fluid in either 5% CO2 in air (20% oxygen; N = 298) or 5% O2, 5% CO2 and 90% N2 (5% oxygen; N = 310) for 7 days. Regardless of the oxygen concentration (5 vs. 20%), rates of pronucleus formation, cleavage, and embryo development up to the eight-cell stage did not differ (46/310 [14.8%] vs. 35/298 [11.7%], respectively; P > 0.05). Moreover, similar proportions of embryos developed in 20 and 5% oxygen tensions (18/298 vs. 27/310). The oocyte nuclear maturation (metaphase II), in terms of decondensed sperm heads, pronucleus formation, cleavage, and embryo development, was similar (P = 0.299) between the atmospheric (20% O2; 12.4% [37/298]) and reduced oxygen tensions (5% O2; 15.8% [49/310]) at all steps of the in vitro culture (IVC) after in vitro fertilization (IVF). To our knowledge, this was the first study that demonstrated that canine embryos can be produced using a low-oxygen in vitro culture system. Both oxygen tensions (5 and 20%) resulted in similar embryonic development and therefore were feasible for IVC of canine oocytes.