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Technology

The aim of the technology was to co-differentiate hiPSCs into cells of all germ layers, such that they can co-develop into a heterogeneous tissue mimicking the process of embryogenesis. Since ectoderm fate is the default pathway for differentiation, a symmetry break was induced by engineering a system to pulse the expression of Gata6, a master transcription factor that guides cell fate towards endoderm and mesoderm lineage. Gata6 was encoded under an inducible promoter and a lentiviral system was used to deliver the transgene in order to achieve heterogeneity in respect to the number of transgene copies received per cell, and consequently, expression level of Gata6. Since a threshold level of Gata6 is required to induce endoderm and mesoderm lineage, the system effectively creates a heterogeneous population of all three germ layers. With sufficient time in culture, paracrine signaling from cell populations results in an organized and heterogeneous tissue.