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A conditionally immortalized cell line from murine proximal tubule

M Loghman-Adham1, A Rohrwasser, C Helin

  • 1Howard Hughes Medical Institute, Department of Human Genetics, University of Utah School of Medicine, Salt Lake City, USA.

Kidney International
|July 1, 1997
PubMed
Summary

Researchers created a conditionally immortalized mouse proximal tubule cell line (tsMPT). These cells differentiate at non-permissive temperatures, offering a valuable model for studying kidney proximal tubule function and differentiation.

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Area of Science:

  • Cell Biology
  • Nephrology
  • Genetics

Background:

  • Developing cell models for kidney research is crucial.
  • Murine proximal tubule cells are essential for studying kidney function.
  • Genetic manipulation capabilities are needed for advanced cell line development.

Purpose of the Study:

  • To develop a conditionally immortalized murine proximal tubule cell line (tsMPT).
  • To establish a cell line suitable for genetic manipulation and differentiation studies.
  • To investigate the functional characteristics of proximal tubule epithelium.

Main Methods:

  • Generation of tsMPT cells from transgenic mice with a temperature-sensitive SV40 large T antigen gene.
  • Culturing cells under permissive (33°C, IFN-γ) and non-permissive (39°C) conditions.

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  • Characterization using immunocytochemistry, Western blotting, electron microscopy, and functional assays.

    • Main Results:

    • tsMPT cells exhibit rapid growth under permissive conditions and differentiate upon transfer to non-permissive conditions.
    • Electron microscopy confirmed cell polarization and apical microvilli.
    • Cells expressed key proximal tubule markers, including brush border enzymes and transporters, and responded to specific stimuli.

    Conclusions:

    • The tsMPT cell line provides a robust in vitro model for proximal tubule research.
    • This model allows for the study of proximal tubule differentiation and function.
    • tsMPT cells are valuable for investigating kidney physiology and disease mechanisms.