Rat embryonic stem-like (ES-like) cells can contribute to extraembryonic tissues in vivo.

Despite significant advances achieved through gene targeting in mouse embryonic stem (ES) cells, this technology is presently only available in mice. Because the rat is a species of undeniable importance to biomedical research, attempts at derivation of rat ES cell lines have been ongoing for many years; however, the putative rat ES cell lines that have been reported to date have not yet displayed the ability to contribute in vivo to developing tissues following embryo injection. In contrast to previous studies, we describe herein the successful derivation and characterization of rat ES-like cell lines that not only express markers of undifferentiated cells, alkaline phosphatase (AP) activity and stage-specific embryonic antigen-1 (SSEA-1) cell surface antigen, but also retain expression of Oct4 (also known as Pou5f1) a homeodomain transcription factor and molecular marker of pluripotent cells. Notably, these rat ES-like cells, when injected into blastocysts transferred to pseudopregnant females, can contribute to developing extraembryonic tissues. This report demonstrates for the first time that rat ES-like cells can be derived efficiently, can express a panel of pluripotent cell markers, can be genetically modified in vitro and cryopreserved, and importantly, are capable of contributing to extraembryonic tissues in vivo.