An In Vitro Method to Analyze Glucose Uptake in Podocytes.

Podocytes are terminally differentiated, insulin-sensitive cells of the glomerular filtration barrier, with a central role in filtration barrier maintenance. Podocyte injury is one of the earliest features observed in diabetic nephropathy (DN) and plays a key role in the development of albuminuria. Several factors are associated with diabetes-mediated podocyte injury, including dysregulated metabolic pathways and insulin sensitivity (Lay & Coward, Nephrol Dial Transplant 29:1127-1133, 2014); thus, reliable assays to study these responses are key in understanding podocyte alterations in DN. Here, we detail an in vitro method to analyze glucose uptake in conditionally immortalized human podocytes (Saleem, J Am Soc Nephrol 13:630-638, 2002; Coward, Diabetes 54:3095-3102, 2005); this assay is useful for detecting changes in podocyte metabolism, nutrient sensing, and insulin sensitivity.