Invited Speaker Presentation Hunter Cell Biology Meeting 2022

Understanding human reprogramming: a journey from epiblast into iBlastoids (#60)

Jose Polo 1
  1. Monash University, Clayton, VIC, Australia

In 2007 Shinya Yamanaka demonstrated that human fibroblasts can be reverted back to a pluripotent state by the forced expression of four transcription factors; OCT4, SOX2, KLF4 and cMYC (OSKM). These so called induced pluripotent stem cells (iPSCs), like embryonic stem cells (ESCs) derived from the epiblast of blastocysts, can give rise to any cell types of the body. Furthermore, iPSCs carry the promise of personalized regenerative medicine and hold tremendous potential for applications such as cell replacements therapeutics, disease modelling and in vitro drug screening. However, he molecular mechanisms of these transitions into primed or naive human-induced pluripotency remains poorly understood. To address this, we reconstructed the molecular reprogramming trajectories using single cell transcriptomics. This revealed that reprogramming into primed and naive human pluripotency follows diverging and distinct trajectories into the pluripotent states. The integration of regulatory element usage with transcriptomics unveiled an unexpected role of trophectoderm (TE) lineage-associated transcription factors as well as a subpopulation of cells that might transiently enter a TE-like state during reprogramming into naive pluripotency. We demonstrated that this transiently upregulated TE state can be stabilised by changing the culture condition, allowing the derivation of induced Trophoblast Stem Cells (iTSCs). Further inspection of this cell cultures revealed also the upregulation of a primitive endoderm like signature in some of the cells. Unexpectedly, when all these cells are allowed to contact each other in a 3D culture, they self-organised giving rise to blastocyst-like structures which we have called iBlastoids. We anticipate that iTSCs and iBlastoids will become pivotal tools in the understanding of human developmental biology, embryology and organ modelling in general since they can provide scalable and tractable  in vitro models of human placentation and blastocysts to study the early steps of human development.