Establishment of a peritoneal mesothelial cell line from a transgenic rat harbouring the temperature-sensitive simian virus 40 large T-antigen gene.

BACKGROUND Establishing a peritoneal mesothelial cell (MC) line in which the native characteristics of primary MCs are constantly maintained in vivo is of great significance for investigating their morphological and functional changes in peritoneal dialysis. We established transgenic (Tg) rats that expressed the temperature-sensitive tsA58 mutant of the simian virus 40 large T-antigen (tsSV40T), which served as a source of immortalized rat cell lines. The cells were immortalized at a permissive temperature of 33 degrees C, although they were differentiated at a non-permissive temperature of 38 degrees C. In this study, we established a novel MC line from tsSV40T Tg rats and evaluated its characteristics. METHODS MCs were isolated from 8-week-old tsSV40T Tg rats and cloned. MCs from 8-week-old Wistar rats were used as controls. These cells were immunohistochemically and phenotypically evaluated by immunofluorescence, phase contrast and electron microscopy. The production of plasminogen activator inhibitor 1 (PAI-1) from MCs stimulated by tumour necrosis factor-alpha (TNF-alpha) was measured. RESULTS The tsSV40T MCs showed a cobblestone-like appearance at 33 and 38 degrees C, which was similar to normal primary cultured MCs. Microvilli-like structures were observed on the cell surface by a scanning electron microscope at 33 and 38 degrees C. Wilms tumour-1 and pancytokeratin, as MC markers, were expressed at 33 and 38 degrees C. Following TNF-alpha stimulation, PAI-1 production of tsSV40T MCs was similar to that of normal primary cultured MCs. CONCLUSION We established a novel, conditionally immortalized MC line that continuously retained the characteristics of primary cultured peritoneal MCs. This cell line might be a useful tool for various types of in vitro biological research on peritoneal dialysis.

[1]  Y. Tomino,et al.  Inhibitory effects of matrix metalloproteinase inhibitor ONO-4817 on morphological alterations in chlorhexidine gluconate-induced peritoneal sclerosis rats. , 2007, Nephrology, dialysis, transplantation : official publication of the European Dialysis and Transplant Association - European Renal Association.

[2]  H. Sugiyama,et al.  Endostatin peptide, an inhibitor of angiogenesis, prevents the progression of peritoneal sclerosis in a mouse experimental model. , 2007, Kidney International.

[3]  M. Jaurand,et al.  Rat pleural mesothelial cells in culture , 1981, In Vitro.

[4]  T. Naito,et al.  Effects of Glucose and Plasminogen Activator Inhibitor‐1 on Collagen Metabolism in the Peritoneum , 2005, Therapeutic apheresis and dialysis : official peer-reviewed journal of the International Society for Apheresis, the Japanese Society for Apheresis, the Japanese Society for Dialysis Therapy.

[5]  Toshikazu Nakamura,et al.  Possible Effects of Hepatocyte Growth Factor for the Prevention of Peritoneal Fibrosis , 2005, Nephron Experimental Nephrology.

[6]  Y. Tomino,et al.  Morphologic changes of peritoneum and expression of VEGF in encapsulated peritoneal sclerosis rat models. , 2004, Kidney international.

[7]  P. Bonniaud,et al.  Basic Mechanisms and Clinical Implications of Peritoneal Fibrosis , 2003, Peritoneal dialysis international : journal of the International Society for Peritoneal Dialysis.

[8]  Y. Hirashima,et al.  Transforming Growth Factor-β1 Produced by Ovarian Cancer Cell Line HRA Stimulates Attachment and Invasion through an Up-regulation of Plasminogen Activator Inhibitor Type-1 in Human Peritoneal Mesothelial Cells* , 2003, Journal of Biological Chemistry.

[9]  J. Nagy,et al.  Mesothelial Cell Transplantation in Models of Acute Inflammation and Chronic Peritoneal Dialysis , 2003, Peritoneal dialysis international : journal of the International Society for Peritoneal Dialysis.

[10]  S. Lowe,et al.  Rb-Mediated Heterochromatin Formation and Silencing of E2F Target Genes during Cellular Senescence , 2003, Cell.

[11]  Misao Suzuki,et al.  Temperature‐dependent, neurotrophic factor‐elicited, neuronal differentiation in adrenal chromaffin cell line immortalized with temperature‐sensitive SV40 T‐antigen , 2003, Journal of neurochemistry.

[12]  Y. Tabuchi,et al.  Development of a conditionally immortalized testicular Sertoli cell line RTS3-3 from adult transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene. , 2003, Cell structure and function.

[13]  J. Jimenez-Heffernan,et al.  Peritoneal dialysis and epithelial-to-mesenchymal transition of mesothelial cells. , 2003, The New England journal of medicine.

[14]  D. Whitaker,et al.  Culturing mouse peritoneal mesothelial cells. , 2003, Pathology, research and practice.

[15]  R. Matusik,et al.  Immortalized epididymal cell lines from transgenic mice overexpressing temperature-sensitive simian virus 40 large T-antigen gene. , 2002, Journal of andrology.

[16]  S. Mutsaers.  Mesothelial cells: Their structure, function and role in serosal repair , 2002, Respirology.

[17]  Y. Tabuchi,et al.  Development and characterization of conditionally immortalized gastric epithelial cell lines from transgenic rats harboring temperature-sensitive simian virus 40 large T-antigen gene. , 2002, Cell structure and function.

[18]  M. Kawahara,et al.  Establishment and characterization of a colonic epithelial cell line MCE301 from transgenic mice harboring temperature-sensitive simian virus 40 large T-antigen gene. , 2000, Cell structure and function.

[19]  H. Ha,et al.  Effect of High Glucose on Peritoneal Mesothelial Cell Biology , 2000, Peritoneal dialysis international : journal of the International Society for Peritoneal Dialysis.

[20]  M. Obinata,et al.  Establishment of SV40-tsA58 transgenic rats as a source of conditionally immortalized cell lines. , 1999, Experimental animals.

[21]  M. Hoopmann,et al.  Intraperitoneal transplantation of isologous mesothelial cells for prevention of adhesions. , 1999, The European journal of surgery = Acta chirurgica.

[22]  R. Zeller,et al.  Rearrangements of the cytoskeleton and cell contacts induce process formation during differentiation of conditionally immortalized mouse podocyte cell lines. , 1997, Experimental cell research.

[23]  B. Banas,et al.  Immortalization and characterization of human peritoneal mesothelial cells. , 1997, Kidney international.

[24]  E. Stylianou,et al.  Isolation, culture and characterization of human peritoneal mesothelial cells. , 1990, Kidney international.

[25]  T. Kooistra,et al.  Characterization and fibrinolytic properties of human omental tissue mesothelial cells. Comparison with endothelial cells. , 1990, Blood.

[26]  M. Jaurand,et al.  Culture procedure of mesothelial cells from the rat parietal pleura. , 1978, Biomedicine / [publiee pour l'A.A.I.C.I.G.].

[27]  Alexandra Eitel.  Senescence , 2014, British medical journal.

[28]  L. Hayflick,et al.  The serial cultivation of human diploid cell strains. , 1961, Experimental cell research.