3D Kidney-like Structures Derived from Rat Kidney Stem/Progenitor Cells

In Vitro Nephrotoxicity Assay Using Three-dimensional Kidney-like Structures Derived from Rat Kidney Stem/Progenitor Cells


Cisplatin damages kidney-like structures prepared by three-dimensional culture of rKS cells.

  • This effect can be extended to assays that evaluate renal impairment and renal regeneration.


We demonstrated the in vitro formation and the course of progressive impairment of kidney-like structures over time.

  • Renal impairment and renal regeneration that occur in the living body were visually represented.

We demonstrated the possibility of constructing three-dimensional structures with high reproducibility.

  • It is possible to provide a stable supply of quality-controlled kidney-like structures.

Visualization of various biomarkers is possible by means of gene transfer.

  • Effective use of biomarkers (mechanism-based evaluation system)

Quantification of renal regeneration and renal impairment (cell death) by imaging analysis.

  • It is expected to standardize instruments for the evaluation of toxicity utilizing the latest microscopes.

The kidney is a complex organ consisting of more than 2 million nephrons and more than 20 types of cells, and it is highly susceptible to drugs. Renal impairment studies are mainly performed by urine tests, blood tests, and renal pathological tests using rats. However, it has become necessary to develop an in vitro nephrotoxicity evaluation method in view of time, cost, and animal welfare. Due to the complexity of kidney structure, it was presumed that the in vitro evaluation of renal function using cultured cells would be djfficult. However, Kitamura, et al., Okayama University, have succeeded in isolating kidney stem/progenitor cells from the proximal tubule S3 segment of rat kidneys and reconstructed kidney-like structures with a nephron structure through three-dimensional culture.


In Vitro Nephrotoxicity Assay Using Three-dimensional Kidney-like Structures Derived from Rat Kidney Stem/Progenitor Cells

Three-dimensional kidney-like structures derived from rKS56 cells were exposed to cisplatin for 6 clays from Day 14 of culture.
They were imaged and analyzed using a three-dimensional cell measurement system (SCREEN Holding Co., Ltd.).
The results demonstrated changes in the number, volume, and, surface area of the structures due to treatment with cisplatin.
We will present the data, including moving images, at the workshop.


In Vitro Nephrotoxicity Assay Using Three-dimensional Kidney-like Structures Derived from Rat Kidney Stem/Progenitor Cells

We aimed to develop a new in vitro nephrotoxicity evaluation method using the said reconstructed three-dimensional kidney-like structures. First, we established a protocol to prepare two or more reconstructed three-climensional kidney-like structures of uniform characteristics at the same time. Next, we chronologically observed the state of reconstruction of three-dimensional kidney-like structures to examine the timing and method of exposure to chemical substances. When reconstructed three-climensional kidney-like structures on Day 14 of culture were treated with cisplatin, the induction of cell death, and the inhibition of the extension of proximal tubule-like structures were observed. If a fluorescent protein is expressed in KS cells, it will be possible to analyze the morphology of three-dimensional kidney-like structures in more detail.

It was possible to establish EGFP-expressing KS cells by using a mouse artificial chromosome vector that was developed at Tottori University. We will discuss the potential of the in vitro nephroroxicity evaluation method using three-dimensional kidney-like structures derived from these KS cells.

This study is the outcome of METI Tox-ln vitro.

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