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Rapid loss of Oct-4 and pluripotency in cultured rodent blastocysts and derivative cell lines.

TitleRapid loss of Oct-4 and pluripotency in cultured rodent blastocysts and derivative cell lines.
Publication TypeJournal Article
Year of Publication2003
AuthorsBuehr M, Nichols J, Stenhouse FH, Mountford P, Greenhalgh CJ, Kantachuvesiri S, Brooker G, Mullins JJ, Smith AG
JournalBiol Reprod
Volume68
Issue1
Pagination222-9
Date Published2003 Jan
ISSN0006-3363
KeywordsAnimals, Animals, Genetically Modified, Base Sequence, Blastocyst, Cell Differentiation, Cell Line, Chimera, Culture Techniques, DNA, Complementary, DNA-Binding Proteins, Gene Expression, In Situ Hybridization, Mice, Mice, Inbred CBA, Octamer Transcription Factor-3, Pluripotent Stem Cells, Rats, Rats, Inbred F344, Rats, Sprague-Dawley, Transcription Factors
Abstract

The POU transcription factor Oct-4 is essential for the pluripotent character of the mouse inner cell mass (ICM) and derivative embryonic stem (ES) cells. We analyzed the expression of Oct-4 during culture and establishment of cell lines from mouse and rat preimplantation embryos. Oct-4 was rapidly lost in primary outgrowths of the majority of cultured embryos prior to any evidence of morphological differentiation. Oct-4 persisted in only a minority of strain 129 cultures, which can go on to give ES cells. We used transgenic rats in which the dual reporter/selection marker beta-geo is under control of Oct-4 regulatory elements to investigate the effect of direct selection for Oct-4 expressing cells. Ablation of all cells occurred, consistent with complete downregulation of Oct-4. Without selection, in contrast, continuous cultures of morphologically undifferentiated cells could be derived readily from rat blastocysts and ICMs. However, these cells did not express significant Oct-4 and, although capable of differentiating into extraembryonic cell types, appeared incapable of producing fetal germ layer derivatives. Downregulation of Oct-4 appears to be a limiting factor in attempts to derive pluripotent cell lines from preimplantation embryos.

Alternate JournalBiol. Reprod.
PubMed ID12493717