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Isolation and propagation in vitro of peritoneal mesothelial cells
J T Hjelle1, B T Golinska, D C Waters
1University of Illinois College of Medicine, Peoria 61656.
Abstract:
Mesothelial cells lining the peritoneal cavity are the primary site of molecular exchange during peritoneal dialysis, a life support system for over 50,000 patients worldwide. In this study, techniques are described for the isolation and propagation in culture of peritoneal mesothelial cells from rats and rabbits. For comparison, mesothelial cells were also obtained from the serosal surface of human colonic tissue. By electron microscopy the cultured cells were found to exhibit microvilli, a well-developed endoplasmic reticulum and golgi apparatus, micropinocytotic vesicles, and lipid-filled intracellular vesicles. Immunochemical probes revealed the expression by these cells in vitro of cytokeratin, fibronectin, vimentin, and keratin, but not von Willebrand factor. Mesothelial cells from rat, rabbit, and human exhibited contact inhibition, but differences in growth rates and dependence on supplements to the growth media. This work provides a multispecies comparison of the behavior of mesothelial cells in vitro for the purpose of developing an experimental system for the study of mesothelial cell biology and the role of these cells in peritoneal dialysis.
Insights
Researchers cultured peritoneal mesothelial cells from rats, rabbits, and humans. This study establishes a multispecies in vitro system to investigate mesothelial cell biology and its role in peritoneal dialysis.
Area of Science:
- Cell Biology
- Biotechnology
- Nephrology
Background:
- Mesothelial cells are crucial for molecular exchange in peritoneal dialysis.
- Peritoneal dialysis is a vital life support system for numerous patients globally.
Purpose of the Study:
- To develop techniques for isolating and culturing rat, rabbit, and human peritoneal mesothelial cells.
- To establish a multispecies in vitro model for studying mesothelial cell behavior.
- To investigate the role of mesothelial cells in peritoneal dialysis.
Main Methods:
- Isolation and propagation of mesothelial cells from rat, rabbit, and human tissues.
- Characterization of cultured cells using electron microscopy.
- Immunochemical analysis to identify protein expression in vitro.
Main Results:
- Cultured mesothelial cells displayed characteristic ultrastructural features and expressed specific proteins (cytokeratin, fibronectin, vimentin, keratin).
- All species' cells exhibited contact inhibition, with variations in growth rates and media supplement needs.
- Absence of von Willebrand factor expression was noted.
Conclusions:
- Successful isolation and culture of multispecies mesothelial cells were achieved.
- The study provides a foundation for an experimental system to study mesothelial cell biology.
- This research aids in understanding the function of mesothelial cells in peritoneal dialysis.