Establishing a co-differentiation culture system from pluripotent stem cells for studying human pancreatic development and disease

Organ development relies on the crosstalk between multiple progenitor cell types. Despite this, most in vitro stem cell models focus solely on differentiating single cell lineages. In this project, we aim to address this gap by establishing a co-differentiation platform to generate pancreatic beta- cells alongside their surrounding mesenchymal cells. By manipulating key developmental pathways from gastrulation, we will simultaneously induce endoderm and mesoderm lineages in the same culture. The resulting populations will be extensively characterised at multiple differentiation stages using qPCR, immunofluorescence and ultimately single-cell RNA sequencing to define lineage composition. Our goal is to obtain beta-cells with enhanced functional maturity while recapitulating the heterogeneity of mesenchymal niches observed in vivo. In the later stages of the PhD, we will leverage this system to investigate epithelial–mesenchymal interactions and explore how these lineages shape one another. We further plan to develop advanced synthetic platforms incorporating microfluidic gradients and synthetic organisers to create artificial gradients and model spatial patterning. Findings from the in vitro systems will be validated using the mouse and mouse pancreatic explants. Collectively, this work will provide new insights into pancreas development, improve our understanding of lineage crosstalk, and inform strategies for beta-cell replacement therapies in diabetic patients.