Bovine ICM derived cells express the Oct4 ortholog.
The goal of this study was to define conditions for the successful isolation of embryonic stem cells from bovine blastocysts. Expression of the Pit-Oct-Unc (POU) transcription factor Oct4 was employed to monitor the pluripotent status of cultured cells. No expression of the previously identified bovine Oct4 pseudogene was found, and transcription of the Oct4 ortholog correlated with the proliferative potential of bovine ICM derived cells. Two methods to isolate pluripotent inner cell mass were compared; 90% of trypsin isolated ICMs formed growing cultures, whereas only 12%-23% of the ICMs isolated by immunosurgery attached and grew. Colony formation from complete blastocysts was 55%. The bovine ICM derived cells could be grown for 4-7 passages. However, Oct4 transcripts were only present in the primary cultures, indicating that the initial culture period of bovine ICM derived cells is critical and needs to be optimized to yield true ES cells. In contrast to bovine ICMs, murine ICMs yielded rapidly growing cells, which proliferated for more than 60 passages.
Yadav
Prem S
PS
Department of Biotechnology, Institute for Animal Breeding (FAL) Mariensee, Neustadt, Germany.
Kues
Wilfried A
WA
Herrmann
Doris
D
Carnwath
Joseph W
JW
Niemann
Heiner
H
D000818:Animals; D001483:Base Sequence; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism*; D002417:Cattle; D049109:Cell Proliferation / Q000187:drug effects; D002469:Cell Separation / Q000379:methods*; D002478:Cells, Cultured; D017124:Conserved Sequence; D004268:DNA-Binding Proteins / Q000235:genetics* / Q000378:metabolism*; D018507:Gene Expression Regulation, Developmental; D051379:Mice; D050814:Octamer Transcription Factor-3; D010641:Phenotype; D039904:Pluripotent Stem Cells / Q000166:cytology / Q000378:metabolism; D011544:Pseudogenes / Q000235:genetics; D017385:Sequence Homology; D014157:Transcription Factors / Q000235:genetics* / Q000378:metabolism*
D004268:DNA-Binding Proteins; D050814:Octamer Transcription Factor-3; C494174:Pou5f1 protein, mouse; D014157:Transcription Factors