Author Spotlight: Investigating Islet Abnormalities and Function with a Pseudoislet Protocol

Our lab is working to understand, prevent, and reverse beta cell dysfunction and other eyelid abnormalities in type one and type two diabetes. Our studies focus on the pathogenesis of human diabetes by integrating studies of the human pancreas and eyelid biology and health and disease. Human eyelets are 3D spherical structures, and accessing cells throughout the entire eyelet presents some experimental challenges.

For example, when trying to introduce biosensors to understand eyelet cell signaling, our pseudo eyelid protocol overcomes this challenge by performing genetic manipulations in the single cell state and aggregating these transduced human eyelet cells into pseudo eyelids for downstream studies. Our pseudo eyelid system allows us to express biosensors throughout the entire eyelet, rather than just the eyelid surface. Combined with our live cell imaging and Microperfusion System, we can both measure and co-register dynamic intracellular processes with downstream hormone secretion.

This protocol could be applied to answer a variety of important scientific questions. For example, gene silencing strategies could be combined with this approach to understand the impact of a gene of interest on eyelid function and associated signaling pathways, either globally or in a self-specific manner.