Monodelphis domestica Induced Pluripotent Stem Cells Reveal Metatherian Pluripotency Architecture.

Marsupials have been a powerful comparative model to understand mammalian biology. However, because of the unique characteristics of their embryology, marsupial pluripotency architecture remains to be fully understood, and nobody has succeeded in developing embryonic stem cells (ESCs) from any marsupial species. We have developed an integration-free iPSC reprogramming method and established validated iPSCs from two inbred strains of a marsupial, The monoiPSCs showed a significant (6181 DE-genes) and highly uniform ( [95% CI] = 0.973 ± 0.007) resetting of the cellular transcriptome and were similar to eutherian ESCs and iPSCs in their overall transcriptomic profiles. However, monoiPSCs showed unique regulatory architecture of the core pluripotency transcription factors and were more like marsupial epiblasts. Our results suggest that and the splice-variant-specific expression of synergistically regulate the opossum pluripotency gene network. It is plausible that , splice variant XM_016427856.1, and form a self-regulatory network. expression, however, was specific to monoiPSCs and epiblasts. Furthermore, was highly expressed in trophectoderm cells, whereas all other pluripotency transcription factors were significantly downregulated, suggesting that the regulatory architecture of core pluripotency genes of marsupials may be distinct from that of eutherians.