Feline embryo development in commercially available human media supplemented with fetal bovine serum

Abstract

Feline embryo development was examined for 7 days after fertilization using commercially available human media supplemented with 0.3% bovine serum albumin (BSA) or 5% fetal bovine serum (FBS). Cumulus-oocyte complexes were categorized as Grades 1, 2, and 3 according to morphology. Only-One Medium (OM) was used for in vitro culture (IVC) in OM + BSA, OM + FBS, and OM + BSA/FBS, with BSA supplementation for the first 2 days and FBS for the subsequent 5 days. Embryos cultured in Early Culture Medium (1–2 days) and Blastocyst Medium (3–7 days) were defined as EB + BSA and EB + BSA/FBS. The developmental rate until the blastocyst stage of Grade 1 and 2 oocytes cultured in OM + BSA/FBS was higher than for the other groups and was significantly higher than for the OM + BSA and EB + BSA groups ( P <0.01). Grade 3 oocytes cultured in OM + BSA/FBS also showed the greatest proportion of blastocyst formation. However, FBS supplementation throughout the IVC period reduced blastocyst number. The percentage of 2 pronuclei after fertilization as well as blastocyst cell number were significantly higher in Grade 1 and 2 than Grade 3 oocytes when cultured in OM + BSA/FBS ( P <0.05). These results indicate that commercially available OM supplemented with BSA for the first 2 days of culture and FBS for the subsequent 5 days is suitable for feline embryo development until the blastocyst stage.

Effects of the preservation medium and storage duration of domestic cat ovaries on the maturational and developmental competence of oocytes <i>in vitro</i>

“…The ovaries of domestic cats (≥5 months) were randomly preserved in physiological saline (Otsuka Pharmaceutical, Tokushima, Japan), EC (Irom Pharmaceutical, Tokyo, Japan), or ET-K (Otsuka Pharmaceutical) for 24, 48, or 72 h and stored at 4 °C until use. The COCs were then collected from fresh and individually stored ovaries and matured in vitro as described by Alam et al [21]. The collected COCs were rinsed three times with IVM medium, which contains Medium 199 (Thermo Fisher Scientific, MA, USA) supplemented with 0.4% (w/v) bovine serum albumin (BSA; Sigma-Aldrich Corp., Mo, USA), 10 IU/mL 17β-estradiol (Sigma-Aldrich), 100 µg/mL gentamycin (Sigma-Aldrich), 137 µg/mL sodium pyruvate (Sigma-Aldrich), 0.02 IU/mL Follistim ® (human recombinant folliclestimulating hormone; MSD, Tokyo, Japan), and 25 ng/mL epidermal growth factor (Sigma-Aldrich).…”