Effects of culture system and protein supplementation on mRNA expression in pre-implantation bovine embryos.
Assisted reproduction technologies have made great progress during the last 15 years in most mammalian species, including humans. Growing evidence indicates that bovine pre-implantation development is a superior model for investigating early human development than the mouse. The purpose of this study was to investigate the effects of two basic culture systems [tissue culture medium (TCM) with 5% CO(2) in air or synthetic oviduct fluid (SOF) with 7% O(2), 88% N(2,) 5% CO(2)] and various protein supplements (serum, bovine serum albumin or polyvinyl alcohol) on the relative abundance of a set of developmentally important gene transcripts in bovine morulae and blastocysts and to compare the results with those for their in-vivo-derived counterparts. The basic culture system including the basic medium composition and oxygen tension had profound effects on the amounts of specific transcripts in bovine embryos, whereas the 'protein source' had only weak effects. Significant differences (P < or = 0.05) in the relative abundance of specific gene transcripts were detected between in-vivo and in-vitro-derived embryos, especially at the morula stage. More differences were found between embryos produced in the TCM system and in-vivo-derived embryos than between SOF-generated embryos and their in-vivo counterparts. No differences were found in the relative abundance of gene transcripts in embryos generated under chemically defined conditions in the two different laboratories. It is concluded that the SOF system provides an environment in which pre-implantation development of bovine embryos is more similar to that occurring in vivo than in the TCM system.
Wrenzycki
C
C
Department of Biotechnology, Institut für Tierzucht und Tierverhalten (FAL), Mariensee, 31535 Neustadt, Germany.
Herrmann
D
D
Keskintepe
L
L
Martins
A
A
Sirisathien
S
S
Brackett
B
B
Niemann
H
H
D000818:Animals; D001769:Blood; D001826:Body Fluids; D002245:Carbon Dioxide / Q000008:administration & dosage; D002417:Cattle; D003470:Culture Media; D046508:Culture Techniques; D004622:Embryo, Mammalian / Q000502:physiology*; D047108:Embryonic Development; D005187:Fallopian Tubes / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D009584:Nitrogen / Q000008:administration & dosage; D010100:Oxygen / Q000008:administration & dosage; D011142:Polyvinyl Alcohol; D011247:Pregnancy; D011506:Proteins / Q000008:administration & dosage*; D012333:RNA, Messenger / Q000032:analysis*; D012710:Serum Albumin, Bovine / Q000008:administration & dosage
D003470:Culture Media; D011506:Proteins; D012333:RNA, Messenger; D002245:Carbon Dioxide; D012710:Serum Albumin, Bovine; D011142:Polyvinyl Alcohol; D009584:Nitrogen; D010100:Oxygen