Amino acid metabolism of preimplantation bovine embryos cultured with bovine serum albumin or polyvinyl alcohol.
Bovine serum albumin (BSA) is an embryotrophic macromolecule used in embryo culture media, which is commonly replaced with synthetic compounds, such as polyvinyl alcohol (PVA). This study compared the effect of BSA and PVA on the development, blastocyst cell number and amino acid metabolism of preimplantation bovine embryos in vitro. Embryos were produced by in vitro maturation and fertilization of immature oocytes from abattoir-derived ovaries. Zygotes were cultured in synthetic oviduct fluid with either 4 mg/ml BSA (SOFaaBSA) or 1 mg/ml PVA (SOFaaPVA) in microdrops with a mineral oil overlay at 39 degrees C under a 5% O2/5% CO2/90% N2 atmosphere. Blastocyst rate and cell numbers were determined after 123 h of culture. In parallel, single expanding blastocysts grown in either medium were incubated in microdrops for 12 h. Amino acid profile of spent drops was determined by high performance liquid chromatography. Replacing BSA with PVA depressed blastocyst rate and cell numbers, and led to quantitative and qualitative differences in amino acid appearance, disappearance and turnover. These differences could partly be due to an increase in free intracellular amino acid concentration in SOFaaBSA embryos derived from hydrolysis of endocytosed BSA, and argue against the inclusion of PVA in bovine embryo culture media.
Orsi
Nicolas M
NM
Department of Biology, The University of York, PO Box 373, York YO10 5YW, UK.
Leese
Henry J
HJ
D000596:Amino Acids / Q000378:metabolism*; D000818:Animals; D001755:Blastocyst / Q000378:metabolism*; D002417:Cattle / Q000196:embryology*; D003470:Culture Media; D046508:Culture Techniques; D047108:Embryonic Development; D005314:Embryonic and Fetal Development; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D011142:Polyvinyl Alcohol; D011247:Pregnancy; D012710:Serum Albumin, Bovine; D015053:Zygote / Q000502:physiology
D000596:Amino Acids; D003470:Culture Media; D012710:Serum Albumin, Bovine; D011142:Polyvinyl Alcohol