Clinical Factors Associated With the Culture of Primary Human Peritoneal Mesothelial Cell From Peritoneal Dialysate Effluent

Background: The culture of primary human peritoneal mesothelial cell (HPMC) provides us an in vitro tool to investigate peritoneal fibrosis and ultrafiltration failure. The aim of the present study was to establish the method of culturing HPMC and to explore clinical factors associated with the success rate of culture.Methods: HPMCs were aseptically harvested by centrifuge from peritoneal dialysate effluent (PDE) of patients with end-stage renal disease (ESRD) who underwent peritoneal dialysis (PD) catheterization for less than 2 weeks and were cultured in vitro. Cells were identified by simple morphological observation and immunofluorescent staining. Clinical data of PD patients was collected. Comparison between groups and binary logistic regression analysis were employed to explore the clinical factors associated with the success rate of culture.Results: The study included 36 patients (26 male (72.2%); mean age 53.9±15.6 years). HPMC from PDE successfully grew and survived in 22 patients. A typical cobblestone-like appearance was observed by inverted phase contrast microscope. Immunofluorescence staining showed positive expression of cytokeratin-18 (CK-18) and Vimentin. Comparison between groups demonstrated significant differences in diabetes (P=0.041), days from catheterization (P=0.002) and the use of erythrocyte lysate (P=0.019) between the two groups. Multivariate logistic regression analysis revealed that the success rate was correlated with days from catheterization (OR=0.318, 95%CI=0.107-0.946, P=0.039) and the level of C-reactive protein (CRP, OR=0.893, 95%CI=0.805-0.991, P=0.032). Conclusions: The method of culturing primary HPMC from PDE has been successfully established. The success rate of culture is correlated with CRP level and days from catheterization.

Involvement of STAT3 Signaling in High Glucose-Induced Epithelial Mesenchymal Transition in Human Peritoneal Mesothelial Cell Line HMrSV5

Background/Aims: Peritoneal fibrosis (PF) is a common complication in patients receiving long-term peritoneal dialysis, which results in damage to peritoneal functions. Epithelial-mesenchymal transition (EMT) is a key step in the early pathogenesis of PF. Increasing evidence has shown that signal transducer and activator of transcription 3 (STAT3) signaling pathway is involved in EMT and tissue fibrosis by interacting with distinct EMT-inducing molecules, including transforming growth factor (TGF)-β and advanced glycation end products (AGEs). This study investigated the involvement of STAT3 in the PF process. Methods: We used high glucose-treated human peritoneal mesothelial cell line HMrSV5 as an in vitro model to expose the peritoneal mesothelial cells to high-glucose dialysate. Expression of EMT markers was detected by qRT-PCR. Accumulation of methylglyoxal (MGO) and AGEs in the culture supernatant were measured by enzyme-linked immunosorbent assay. Phosphorylation of STAT3 was assessed by Western blot. Results: Results showed that high glucose upregulated TGF-β, increased the productions of MGO and AGEs, and induced EMT in HMrSV5 cells. High glucose also activated the STAT3 pathway. STAT3 inhibitor reduced the high glucose-induced EMT, via reducing TGF-β expression and repressing the accumulation of MGO and AGEs. Conclusion: Our results revealed a critical role for STAT3 signaling in high glucose-induced EMT in HMrSV5 cells, and suggested that inhibition of STAT3 might be a treatment for high glucose-induced fibrogenesis in PF.