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Embryo |
a Centre for Genome Research, University of Edinburgh, Edinburgh EH9 3JQ, United Kingdom
b Stem Cell Sciences Ltd., Elsternwick, Victoria 3185, Australia
c Walter and Eliza Hall Institute, Royal Melbourne Hospital, Melbourne, Victoria 3050, Australia
d Medical Radiology, University of Edinburgh, Edinburgh EH8 9AG, United Kingdom
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 ß-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.
2 Correspondence: Mia Buehr, Centre for Genome Research, University of Edinburgh, King's Buildings, West Mains Rd., Edinburgh EH9 3JQ, U.K. FAX: 44 131 650 3777; mbuehr{at}srv0.bio.ed.ac.uk
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