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Optimized Method for Establishing Primary Human Mesothelial Cell Cultures Preserving Epithelial Phenotype.

Evdokiya Kuznetsova1, Nadezhda Bakalenko1, Liana Gaifullina1

  • 1Institute of Cytology, Russian Academy of Sciences, 194064 St-Petersburg, Russia.

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|December 30, 2025
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Summary

Researchers developed a new method to culture human mesothelial cells (MCs), preventing their transition to a mesenchymal state. This breakthrough supports studying serosal diseases and testing therapies.

Keywords:
in vitro cell modelsmesothelial cells (MCs)mesothelial-to-mesenchymal transition (MMT)primary cell cultures

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Area of Science:

  • Cell Biology
  • Pathology
  • Regenerative Medicine

Background:

  • Mesothelial cells (MCs) are crucial for understanding serosal diseases, fibrosis, inflammation, and cancer.
  • Maintaining the epithelial phenotype of MCs in vitro is challenging due to mesothelial-to-mesenchymal transition (MMT).

Purpose of the Study:

  • To establish a reliable method for culturing primary human mesothelial cells.
  • To overcome the challenge of mesothelial-to-mesenchymal transition (MMT) in vitro.
  • To enable functional studies and therapeutic assessments using MCs.

Main Methods:

  • A combined protocol using collagen IV coating.
  • Inclusion of conditioned medium in cell culture.
  • Short-term treatment with a ROCK inhibitor.

Main Results:

  • Improved survival of mesothelial cells in primary cultures.
  • Successful maintenance of the epithelial-like phenotype of MCs.
  • Established a robust method for MC culture.

Conclusions:

  • The developed protocol effectively preserves mesothelial cell phenotype in vitro.
  • This method facilitates research into mesothelial cell function and disease pathogenesis.
  • It provides a valuable platform for evaluating potential therapeutic strategies for serosal diseases.