Shaking culture improves physiological maintenance of primary rat kidney tissue slices

Maintaining viability and physiological function in kidney tissue slice cultures remains a major challenge for ex vivo renal models, primarily due to limited oxygen diffusion into the tissue interior. In this study, we investigated whether shaking culture could improve the maintenance of primary rat kidney tissue slices.

Rat kidney tissue slices (200 µm) were cultured under static or shaking conditions for up to 7 days. Tissue viability, histological features, renal transporter activity, nephrotoxic responses, and metabolomic profiles were evaluated.

Shaking culture markedly improved tissue viability, as evidenced by sustained ATP levels over 7 days compared with static conditions. Histological analysis revealed partial preservation of epithelial polarity, indicated by the maintained apical localization of aquaporin 1. Transporter studies demonstrated that uptake activities of Oat1/3 and Oct2 substrates were better preserved under shaking conditions. Furthermore, shaking-cultured slices enabled the evaluation of nephron segment-specific injury, with cisplatin inducing tubular damage and puromycin or dasatinib affecting glomerular structures. Metabolomic analysis revealed distinct alterations in central carbon metabolism, including increased levels of tricarboxylic acid cycle intermediates, suggesting improved mitochondrial function.

These findings demonstrate that shaking culture provides a simple and effective approach to improve the culture microenvironment, thereby improving the maintenance of viability and physiological function in kidney tissue slices. This system may serve as a useful platform for investigating renal pharmacokinetics and drug-induced kidney injury.