Determination of innervation in pancreas organids co-cultured with neural crest cells

Although, the pancreas is an organ intensely innervated by sympathetic and parasympathetic nerve fibers, pancreatic organoids consist only of epithelial cell derivatives. In pancreatic cancer, there is a bidirectional relationship between neurons and non-neural pancreatic cells characterized by neural plasticity. In this thesis, it was first aimed to create "innervated pancreatic organoids" and to examine pancreatic neuropathy in these organoids. For this purpose, pancreatic organoids were isolated from normal C57BL/6 and Kras+/LSLG12D Trp53fl/fl p48+/Cre (KPC) mice, and these organoids were respctively co-cultured with human induced pluripotent stem cells (hiPSC) derived neural crest cells (NCCs) and dorsal root ganglion (DRG) isolated from C57BL/6 neonatal mice. It was observed that NCCs co-cultured with pancreatic organoids exhibited neuronal morphology and neuronal extensions of DRGs co-cultured with pancreatic organoids began to emerge on the 3rd day of co-culturing. The co-cultures were terminated on the 7th day and the samples were labeled with Anti-E-cadherin, Anti-β3-Tubulin and Anti-GFAP antibodies with the wholemount IF method and visualized by confocal microscopy. In both co-culture models, it was observed that β3-Tubulin positive neurons developed and their extensions directed towards E-cadherin positive organoids and partially interacted. When the developed innervated pancreatic organoids were examined in terms of pancreatic neuropathy, it was determined that KPC pancreatic organoids increased neural plasticity in innervated organoid models formed as a result of both NCCs and DRG co-culture. In this study, it was shown for the first time in the literature that neurons derived from NCCs or mature neurons from DRG can lead to innervation of pancreatic organoids and result in modulation of neuro-exocrine cell interaction in the pancreatic cancer microenvironment.