Inhibitory activity of dome formation in LLC-PK1 cells is a selective index of aminoglycoside nephrotoxicity.

Aminoglycoside antibiotics are widely used clinically for a treatment of Gram-negative bacterial infection by the inhibition of bacterial protein synthesis. However, the aminoglycosides have clinical side effects such as nephrotoxicity and ototoxicity for a long-term treatment. Here we evaluated a simple and useful method for screening a new aminoglycoside without or less nephrotoxicity using LLC-PK1 cells derived from a pig kidney proximal tubular epithelial cell. Dome formation is a morphological trait of LLC-PK1 cells through unidirectional water and salt transport in normal culture condition1,2). In the course of searching for a practical method for evaluating aminoglycosides nephrotoxicity, we reconfirmed usefulness of dome formation in LLC-PK1 cells, which was originally reported by INUI et al.1,2). In this study, we modified and qualified the method for screening a new aminoglycosides, and determined whether it is applied to other antibiotics. As a result, an optimal condition was determined as follows. LLC-PK1 cells obtained from American Type Culture Collection were maintained in a Dulbecco's modified Eagle's medium (DMEM; Nissui Pharmaceutical Co., Ltd.) supplemented with 10% fetal bovine serum