F Y Liu1, S B Duan, Z G Long. 1. Department of Nephrology, Second Xiangya Hospital, Central South University, Chiangsha 410011, China.
Abstract
OBJECTIVE: To establish a reproducible model for culture of human peritoneal mesothelial cells (HPMCs), and observe the expressions of extracellular matrix (ECM) protein and interleukin-8 (IL-8) in HPMC. METHODS: Mesothelial cells were isolated from human omenta by trypsin EDTA disaggregation. HPMCs were identified by morphology and streptomyces protein-peroxidase (SP) method. In the same way, expressions of fibronectin, collagen type III, V and transforming growth factor beta 1 (TGF beta 1) were measured. Expression of IL-8 mRNA and FN mRNA in the HPMCs was detected by reverse transcription (RT) and polymerase chain reaction (PCR) amplification. RESULTS: Confluent HPMCs appeared multipolar under microscope; numerous surface microvilli and an abundant endoplasmic reticulum were observed. Cultured HPMCs coexpressed cytokeratin and vimentin, and synthesized fibronectin, TGF beta 1 and collagen types III (instead of type V). Expressions of IL-8 mRNA and FN mRNA were also observed in HPMCs. CONCLUSION: Establishment of cultured HPMCs model will provide the basis of the research in preventment of peritoneal fibrosis and roles of IL-8 in peritoneal dialysis.
OBJECTIVE: To establish a reproducible model for culture of human peritoneal mesothelial cells (HPMCs), and observe the expressions of extracellular matrix (ECM) protein and interleukin-8 (IL-8) in HPMC. METHODS: Mesothelial cells were isolated from human omenta by trypsin EDTA disaggregation. HPMCs were identified by morphology and streptomyces protein-peroxidase (SP) method. In the same way, expressions of fibronectin, collagen type III, V and transforming growth factor beta 1 (TGF beta 1) were measured. Expression of IL-8 mRNA and FN mRNA in the HPMCs was detected by reverse transcription (RT) and polymerase chain reaction (PCR) amplification. RESULTS: Confluent HPMCs appeared multipolar under microscope; numerous surface microvilli and an abundant endoplasmic reticulum were observed. Cultured HPMCs coexpressed cytokeratin and vimentin, and synthesized fibronectin, TGF beta 1 and collagen types III (instead of type V). Expressions of IL-8 mRNA and FN mRNA were also observed in HPMCs. CONCLUSION: Establishment of cultured HPMCs model will provide the basis of the research in preventment of peritoneal fibrosis and roles of IL-8 in peritoneal dialysis.
Authors: Myriam Holl; Lucas Becker; Anna-Lena Keller; Nora Feuerer; Julia Marzi; Daniel A Carvajal Berrio; Peter Jakubowski; Felix Neis; Jan Pauluschke-Fröhlich; Sara Y Brucker; Katja Schenke-Layland; Bernhard Krämer; Martin Weiss Journal: Biomedicines Date: 2021-02-10