Kidney Week

Abstract: TH-PO0263

Endothelial Coculture Attenuates Flow-Induced Mesangial Activation in a Three-Dimensional (3D) Collagen-Based Culture System

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

• Yoshihara, Maki, Saga Daigaku Igakubu Fuzoku Byoin, Saga, Saga Prefecture, Japan

Maki Yoshihara, MD

• Nishiyama, Megumi, Saga Daigaku Igakubu Daigakuin Igakukei Kenkyuka, Saga, Saga Prefecture, Japan

Megumi Nishiyama, MD, PhD

• Fukuda, Makoto, Saga Daigaku Igakubu Fuzoku Byoin, Saga, Saga Prefecture, Japan

Makoto Fukuda, MD, PhD

• Iwamoto, Shuhei, Saga Daigaku Igakubu Daigakuin Igakukei Kenkyuka, Saga, Saga Prefecture, Japan

Shuhei Iwamoto, PhD

• Sakumoto, Takehisa, Saga Daigaku Igakubu Daigakuin Igakukei Kenkyuka, Saga, Saga Prefecture, Japan

Takehisa Sakumoto, DrMed

• Yumeka, Mine, Saga Daigaku Igakubu Daigakuin Igakukei Kenkyuka, Saga, Saga Prefecture, Japan

Mine Yumeka, MSc

• Yamasaki, Masatora, Saga Daigaku Igakubu Fuzoku Byoin, Saga, Saga Prefecture, Japan

Masatora Yamasaki, MD, PhD

• Kawasaki, Maki, Saga Daigaku Igakubu Daigakuin Igakukei Kenkyuka, Saga, Saga Prefecture, Japan

Maki Kawasaki, MD, PhD

• Miyazono, Motoaki, Saga Daigaku Igakubu Fuzoku Byoin, Saga, Saga Prefecture, Japan

Motoaki Miyazono, MD, PhD

• Aoki, Shigehisa, Saga Daigaku Igakubu Daigakuin Igakukei Kenkyuka, Saga, Saga Prefecture, Japan

Shigehisa Aoki, MD

Group or Team Name

• Division of Nephrology, Department of Internal Medicine, Faculty of Medicine

Background

Mesangial hypercellularity and matrix accumulation are key lesions of mesangial proliferative kidney diseases. Most in vitro systems do not incorporate 3D matrix architecture, mesangial–endothelial interaction, and mechanical cues. We developed a 3D collagen-based coculture system to examine flow-like stimulation and endothelial coculture.

Methods

Mouse mesangial cells (MES13) were embedded within type I collagen gels, and mouse microvascular endothelial cells (MS-1) were seeded onto the apical surface. MES13 monocultures served as controls. Collagen discs were maintained for 7 days under static or fluid-flow conditions using an orbital shaker at 50 rpm. Biomechanical analysis was performed. Solute transport was evaluated by indigo carmine intensity using ImageJ. Cellular responses were evaluated by Ki-67, cleaved caspase-3, Picrosirius Red, type III collagen, and immunoblotting for p38 MAPK, AKT, and ERK phosphorylation.

Results

Indigo carmine intensity decreased over time under both conditions, with a greater decrease under fluid-flow conditions up to 240 min. Under fluid flow, mesangial cell proliferation was accelerated in monoculture, whereas endothelial coculture attenuated this proliferation with reduced Ki-67 labeling. Cleaved caspase-3 labeling was also reduced with endothelial cells. Endothelial coculture suppressed mesangial p38 MAPK phosphorylation and attenuated flow-induced AKT phosphorylation. Fluid flow enhanced Picrosirius Red–positive collagen depositio n and pericellular type III collagen immunoreactivity in mesangial monoculture; these responses were reduced by endothelial coculture.

Conclusion

Orbital shaker–induced flow-like stimulation promoted mesangial proliferation and collagen matrix accumulation. Endothelial coculture attenuated these responses and suppressed mesangial p38 MAPK and AKT phosphorylation.

Acknowledgment

AI disclosure: Claude Opus 4, developed by Anthropic and accessed via claude.ai on May 12, 2026, assisted with complementary biomechanical analysis and numerical calculations based on experimental parameters provided by the authors. All outputs were reviewed and verified by the authors, who take full responsibility for the submitted content.

Funding

• Government Support – Non-U.S.