Isolation and Characterization of Renal Tubular Epithelial Cells

The other portion of kidney tissue was used for isolation of RTECs by a method described previously, with slight modifications [19 (link), 20 (link)]. Briefly, the cortex was cut into fragments. Cells were dissociated by incubation for 30 min at 37°C with 1 mg/mL type-I collagenase. Red blood cells were removed by lysis. RTECs were separated by Percoll gradient density centrifugation [21 (link)]. Purity of RTECs was examined by immunostaining with cytokeratin-18 and Hoechst dye [22 (link)] (Supplementary Figure 2). Isolated cells were used for detection of mitochondrial function (mitochondrial membrane potential (ΔΨm), cellular level of adenosine triphosphate (ATP), mitochondrial permeability transition pore (mPTP), and lysosomal stability), as described previously [15 (link)].