Isolation of functional dendritic cells from murine kidneys for immunological characterization

ABSTRACT Aim: The kidney is a complex organ, requiring the contributions of multiple cell types to perform its various functions. Within this system the dendritic cell has been demonstrated to play a key role in maintaining the immunological balance of the kidney. In this methods paper we aim to identify the best method for isolating murine renal dendritic cells. Methods: The efficiency of isolating dendritic cells from enzymatically digested renal parenchyma by density centrifugation, MACS and FACS was compared. Results: Density centrifugation enriched dendritic cells by only approximately two fold. However, MACS and FACS resulted in a much higher purity (80% versus 95% respectively). Conclusions: Although FACS gave the highest purity, MACS is the optimal method for isolating dendritic cells given cost and time factors. Isolation of a homogeneous population of renal dendritic cells will enable the molecular and functional dissection of these cells in both homeostasis and disease models. This article provides a hands‐on guide to the isolation of dendritic cells from the mouse kidney, a technique that is becoming widely used in the study of immunologic kidney disease. In addition, these methods can be easily adapted for the isolation of other leukocyte populations from the mouse or rat kidney.