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Establishment of renal proximal tubule cell lines derived from the kidney of p53 knockout mice

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Abstract

The human cell line HK-2 is most commonly used as a model of renal proximal tubular epithelial cells (PTECs) for various studies despite the absence or low expression of transporters characteristic of parental PTECs. In an effort to develop reliable PTEC models, several human cell lines have been newly established over the last decade. In contrast, reliable mouse PTEC models are still unavailable. In this study, we established immortalized renal cortex tubule cell lines derived from p53 knockout mice and evaluated various PTEC characteristics toward the development of reliable mouse PTEC models. Here, we focus on MuRTE61, one of 13 newly established clonal cell lines. Albumin uptake in MuRTE61 cells was verified by incubation with fluorescent dye-labeled albumin. RT-PCR confirmed the expression of efflux transporter genes characteristic of PTECs in the MuRTE61 cells. MuRTE61 cells exhibited high sensitivity to treatment with cisplatin, a nephrotoxic agent, accompanied by upregulated expression of the uptake transporter Slc22a2 gene. Furthermore, MuRTE61 cells consistently formed spheroids with a lumen and apicobasal polarity in three-dimensional Matrigel cultures. Apical brush border microvilli were also observed in the spheroids by transmission electron microscopy. These data validate that MuRTE61 can be characterized as a reliable mouse PTEC line. In future, detailed analysis of reliable mouse and human PTEC lines will provide an accurate extrapolation of results of experiments using mice and humans, and may help resolve apparent inconsistencies between mouse and human nephrotoxicity.

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Funding

This work was supported by the Ministry of Education, Culture, Sports, Science and Technology (MEXT), Glant-in-Aid for Scientific Research, KAKENHI (16K16606) to HS and KAKENHI (25430083) to NS. The funders had no role in the design of the study and collection, analysis, decision to publish, interpretation of data or preparation of the manuscript.

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  1. Laboratory of Laboratory Animal Science and Medicine, School of Veterinary Medicine, Kitasato University, 35-1, Higashi-23, Towada, Aomori, 034-8628, Japan

    Hayato Sasaki & Nobuya Sasaki

  2. Laboratory of Veterinary Anatomy, School of Veterinary Medicine, Kitasato University, Towada, 034-8628, Japan

    Makoto Sugiyama

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  1. Hayato Sasaki
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Correspondence to Hayato Sasaki.

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All research was conducted according to the Regulation for the Care and Use of Laboratory Animals of Kitasato University. Animal experimentation protocol was approved by President of Kitasato University through the judgment by Institutional Animal Care and Use Committee of Kitasato University (Approval ID: No. 15-168).

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Sasaki, H., Sugiyama, M. & Sasaki, N. Establishment of renal proximal tubule cell lines derived from the kidney of p53 knockout mice. Cytotechnology 71, 45–56 (2019). https://doi.org/10.1007/s10616-018-0261-1

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