Human pluripotent stem cell-derived acinar/ductal organoids generate human pancreas upon orthotopic transplantation and allow disease modelling

Abstract

ObjectiveThe generation of acinar and ductal cells from human pluripotent stem cells (PSCs) is a poorly studied process, although various diseases arise from this compartment.DesignWe designed a straightforward approach to direct human PSCs towards pancreatic organoids resembling acinar and ductal progeny.ResultsExtensive phenotyping of the organoids not only shows the appropriate marker profile but also ultrastructural, global gene expression and functional hallmarks of the human pancreas in the dish. Upon orthotopic transplantation into immunodeficient mice, these organoids form normal pancreatic ducts and acinar tissue resembling fetal human pancreas without evidence of tumour formation or transformation. Finally, we implemented this unique phenotyping tool as a model to study the pancreatic facets of cystic fibrosis (CF). For the first time, we provide evidence that in vitro, but also in our xenograft transplantation assay, pancreatic commitment occurs generally unhindered in CF. Importantly, cystic fibrosis transmembrane conductance regulator (CFTR) activation in mutated pancreatic organoids not only mirrors the CF phenotype in functional assays but also at a global expression level. We also conducted a scalable proof-of-concept screen in CF pancreatic organoids using a set of CFTR correctors and activators, and established an mRNA-mediated gene therapy approach in CF organoids.ConclusionsTaken together, our platform provides novel opportunities to model pancreatic disease and development, screen for disease-rescuing agents and to test therapeutic procedures.

Engineering tissue‐specific blood vessels

“…Several disease models have been developed using iPSCs derived from patients. These include, but are not limited to, Alzheimer's disease, cystic fibrosis in the pancreas, and gastric Helicobacter pylori infection models using self‐assembled iPSC‐derived tissues. As our understanding of tissue specific vasculature increases, the methods described in this article to engineer vasculature for organ models can be further adapted to incorporate vasculature in the organ‐specific disease models to elucidate vascular mechanisms, discover drug candidates, and examine systemic drug delivery.…”

Using 3D Organoid Cultures to Model Intestinal Physiology and Colorectal Cancer

“…While phenotypes of epithelial-derived cultures from these patients have not yet been reported, intestinal organoids differentiated from hair follicle stem cells of CD patients demonstrate no differences in morphology, perhaps due to lack of immune-derived contributions [60]. However, useful insights have still been made for inflammatory diseases.…”

Take a deep breath and digest the material: organoids and biomaterials of the respiratory and digestive systems

“…[75] Pancreatic organoids have been derived from primary human pancreatic ductal cells and hPSCs by using Matrigel to provide a three-dimensional environment. [76][77][78] Pancreatic organoids derived from the primary ductal cells only contain ductal cells with no acinar or endocrine cells. Even when non-ductal cells were mixed with the ductal cell population, the ductal cell population out-competed the nonductal cells within the organoid cultures.…”

“…After orthotropic transplantation of the pancreatic organoids, ductal and acinar cells were observed with a small population of endocrine cells, resembling both transcriptionally and structurally the fetal pancreas. [78] The challenge thus far is to derive a pancreatic organoid that possesses exocrine and endocrine cell populations. This is particularly challenging due to the nature of the exocrine compartment, which produces a significant number of digestive enzymes and proteases.…”