A Human Pancreatic Spheroid Platform for Toxicology and Diabetes Research

Diabetes mellitus (DM) is a prevalent and chronic metabolic disorder that affects more than 300 million patients in the world. One of the major obstacles in DM is the lack of in vitro models that can capture the physiological features of the in vivo environment. Therefore, to identify new treatments for DM there is an increasing demand for functional in vitro pancreatic islet models in which the cellular microenvironment, consistency and long-term culture is more fully preserved. More recently, functional beta-cells derived from human induced pluripotent stem cells (hiPSCs) have gained prominence as an enticing system for the study of DM as a human, tissue-specific model to model the disease and identify potential therapeutic intervention. Here we describe and characterize a novel micro-engineered 384 well plate platform in which each well contains a single size-controlled hiPSC-pancreatic islet equivalent spheroid that emulates key characteristics of adult human pancreas. Gene expression and immunofluorescence analysis indicated that these islet-equivalents express key markers of pancreas cellular identity such as the master pancreatic transcription factor PDX-1, glucose-sensing and insulin-secreting genes such as GLUT2, ABCC8 and SYT4, and importantly, secreted proteins produced in mature humans islets, like Insulin, glucagon and somatostatin. RNAseq data indicated larger transcriptional consistency from different batches of microPancreas production than traditional cadaveric islet samples. In addition, cells show glucose dependent stimulated insulin secretion (GSIS), highlighting the functionality of the platform. Altogether, we describe a novel human pancreatic platform applicable to gene-expression, imaging and insulin-secretion studies as a reliable research source for diabetes drug discovery, disease modeling and toxicology applications. Disclosure S. Mora-Castilla: Employee; Self; StemoniX. B. Gaertner: Employee; Self; StemoniX. I. Barbosa-Farias: Employee; Self; StemoniX. W. Si: Employee; Self; StemoniX. F. Zanella: Employee; Self; StemoniX.