MethodsX (Jan 2023)
Insulin secretion assays in an engineered MIN6 cell line
Abstract
Insulin secretion from pancreatic beta cells is crucial for maintaining glucose homeostasis. The murine insulinoma derived MIN6 cell line is commonly used as a model for insulin secretion studies. However, its glucose responsiveness wanes with passaging, and insulin secretion is traditionally measured by expensive and time-consuming RIA or ELISA. We have developed a MIN6 subclone (MIN6-6) that allows for high throughput assay of insulin secretion in both population and single cells. In addition, MIN6-6 also expresses Cas9, permitting genome wide CRISPR screen of insulin secretion using a pooled sgRNA library. Here we provide methods for assaying insulin secretion both in bulk and in single cells in MIN6-6 cells, as well as for CRISPR screen of insulin secretion. • A highly glucose responsive beta cell reporter line (MIN6-6) with multiple engineered functionalities. • Allows for CRISPR/Cas9 mutagenesis, quantification of bulk insulin secretion by a straightforward nanoLuc assay and visualization of intracellular insulin granules. • Allows for en masse quantification of insulin granule exocytosis in individual cells under multiple conditions.
