[A modified method for isolating and culturing peritoneal mesothelial cells in rats].

OBJECTIVE

To establish a culture model of peritoneal mesothelial cells in vitro separated by infusing the digestive fluid in the rat abdominal cativity.

METHODS

Peritoneal mesothelial cells (PMC) were digested with 0.125% trypsin-0.01% EDTA Na2 in the abdominal cavity. We collected coelio-perfusate and cultured it. The isolated cells were identified by the phase contrast microscope, electron microscope and immunohistochemical analysis.

RESULTS

The confluent cells showed a uniform cobblestone-like appearance under the phase contrast microscope; numerous surface microvilli could be found under the electron microscope and the purity of the cells was over 95%. Immunohistochemical studies revealed a positive staining for cytokeratin and vimentin, but a negative staining for Factor VIII associated antigen and CD45.

CONCLUSION

The model of rat peritoneal mesothelial cells has been established successfully and it will provide an experimental basis of studying the fibrosis in peritoneal dialysis.