Effects of bovine oviduct epithelial cells, fetal calf serum and bovine serum albumin on gene expression in single bovine embryos produced in the synthetic oviduct fluid culture system.
In this study the synthetic oviduct fluid (SOF) system with bovine oviduct epithelial cell (BOEC) co-culture is compared with an SOF system with common protein supplements. One thousand six hundred bovine embryos were cultured in SOF media supplemented with BOEC, fetal calf serum (FCS) and bovine serum albumin (BSA). Eight different culture groups were assigned according to the different supplementation factors. Developmental competence and the expression levels of five genes, namely glucose transporter-1 (Glut-1), heat shock protein 70 (HSP), connexin43 (Cx43), (2)-actin (ACTB) and glyceraldehyde-3-phosphate dehydrogenase (GAPDH), analysed as mRNA by using reverse transcription-polymerase chain reaction, were measured on bovine embryos cultured for 9 days. Gene expression of these in vitro-produced embryos was compared with the gene expression of in vivo-produced embryos. There was no significant difference found in embryo developmental competence between the Day 9 embryos in BOEC co-culture, FCS and BSA supplements in SOF media. However, differences in gene expression were observed. With respect to gene expression in in vivo and in vitro embryos, BOEC co-culture affected the same genes as did supplementation with FCS and BSA. HSP was the only gene that differed significantly between in vitro and in vivo embryos. When the different in vitro groups were compared, a significant difference between the BOEC co-culture and the FCS supplementation groups due to Glut-1 expression was observed.
Norwegian School of Veterinary Science, 0033 Oslo, Norway.
Mona E
ME
Pedersen
Øzen Banu
ØB
Øzdas
Wenche
W
Farstad
Aage
A
Tverdal
Ingrid
I
Olsaker
D000199:Actins / Q000378:metabolism; D000818:Animals; D002417:Cattle; D018031:Connexin 43 / Q000378:metabolism; D017931:DNA Primers; D046149:Embryo Culture Techniques / Q000379:methods* / Q000662:veterinary*; D004622:Embryo, Mammalian / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology*; D005187:Fallopian Tubes / Q000166:cytology*; D005260:Female; D005312:Fetal Blood / Q000378:metabolism*; D015870:Gene Expression; D051272:Glucose Transporter Type 1 / Q000378:metabolism; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases / Q000378:metabolism; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012710:Serum Albumin, Bovine / Q000378:metabolism*
D000199:Actins; D018031:Connexin 43; D017931:DNA Primers; D051272:Glucose Transporter Type 1; D018840:HSP70 Heat-Shock Proteins; D012710:Serum Albumin, Bovine; D005987:Glyceraldehyde-3-Phosphate Dehydrogenases