Abstract: TH-PO0276
Landscape of Epithelial-Mesenchymal Transition Markers in Co-Cultured Kidney Cells
Session Information
- Glomerular Diseases: Cell Biology
October 22, 2026 | Location: Exhibit Hall A, Convention Center
Abstract Time: 10:00 AM - 12:00 PM
Category: Glomerular Diseases
- 1401 Glomerular Diseases: Mechanisms, including Podocyte Biology
Authors
- Prabhakar, Sharma S., Texas Tech University Health Sciences Center School of Medicine, Lubbock, Texas, United States
- Chatterjee, Arunita, Texas Tech University Health Sciences Center School of Medicine, Lubbock, Texas, United States
Background
Studying monocultured cells simplifies the complexities inherent in biology, allowing researchers to test hypotheses. However many cellular events occur in close crosstalk with other cells in health and disease. . Here, we hypothesized that co-culturing mesangial cells (RMCs) and proximal tubular epithelial cells (TKPTS)—would provide better insights in the context of diabetic micro-environment than monocultures.
Methods
Equal number of cells was seeded into monoculture or coculture flask in DMEM (5.5mM glucose, LG) supplemented with 10% FBS and incubated at 37°C with 5% CO2. Since RMCs have a slower growth rate than TKPTS, the cocultures were seeded at a 2:1 ratio, i.e., twice as many RMCs as TKPTS. The test plates were supplemented after 2 hours with glucose (25mM, HG) to simulate a diabetic milieu. After 48h, cells were isolated, total protein extracted and quantified. Changes in protein expression were assessed by Western blotting.
Results
Co-cultured cells had a high basal expression of EMT markers, which decreased with HG. Expression of β-catenin, an EMT marker and a fibrotic inducer, increased under HG in cultured RMCs and TKPTS.. Interestingly, co-cultured cells had higher β-catenin expression in LG than in HG. A similar pattern occured in the expression of vimentin and claudin 1. Vimentin is strongly expressed in RMC and increased with HG. unlike in TKPTS. while co-cultured cells, its expression was lower in HG than in LG. Claudin 1 was expressed in TKPTS.equally in LG and HG.while in co-cultured cells expressed higher claudin 1 in LG than in HG.
Conclusion
Co-cultured MCs and PTECs induced an EMT-like state in each other under LG conditions, but coped better with HG than monocultures. The proportions of each cell type in the co-culture extracts vary with media glucose levels.. In diabetes, while serum glucose levels are very high, intracellular glucose levels may be low. Compared to single-cell cultures in HG, co-cultured renal cells in LG conditions might provide better insights into pathogenesis of DKD.
Acknowledgment
We sincerely acknowledge the Woihaye Renal Research Endowment for supporting the research work cited here.
Funding
- Private Foundation Support