Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development

作者： Joseph S. Uzarski1,2, Jimmy Su1,2,3,4, Yan Xie1,2, Zheng J. Zhang1,2, Heather H. Ward5, Angela Wandinger-Ness6, William M. Miller7,8, Jason A. Wertheim1,2,3,4,8,9 1Comprehensive Transplant Center, Feinberg School of Medicine,Northwestern University, 2Department of Surgery, Feinberg School of Medicine,Northwestern University, 3Department of Biomedical Engineering,Northwestern University, 4Simpson Querrey Institute for BioNanotechnology in Medicine,Northwestern University, 5Department of Internal Medicine,University of New Mexico HSC, 6Department of Pathology,University of New Mexico HSC, 7Department of Chemical and Biological Engineering,Northwestern University, 8Chemistry of Life Processes Institute,Northwestern University, 9Department of Surgery,Jesse Brown VA Medical Center

简介：

Overview

This protocol outlines the decellularization of Sprague Dawley rat kidneys through antegrade perfusion of detergents, resulting in acellular renal extracellular matrices. These matrices serve as templates for repopulation with human renal epithelial cells, monitored using a resazurin perfusion assay within specialized bioreactors.

Key Study Components

Area of Science

• Nephrology
• Tissue Engineering
• Cell Biology

Background

• Decellularization is a technique used to create scaffolds for tissue engineering.
• Human renal epithelial cells can potentially restore kidney function.
• Perfusion bioreactors provide a controlled environment for cell growth.
• Monitoring cell viability is crucial for assessing scaffold repopulation.

Purpose of Study

• To develop a method for repopulating acellular kidney scaffolds with human cells.
• To evaluate the effectiveness of the repopulation process.
• To investigate factors influencing cell growth and differentiation.

Methods Used

• Decellularization of rat kidneys using detergent solutions.
• Injection of human renal cortical tubular epithelial cells into the scaffolds.
• Continuous antegrade arterial perfusion within a bioreactor system.
• Assessment of cell growth using resazurin perfusion assay and immunohistochemical analysis.

Main Results

• Successful decellularization resulted in viable extracellular matrix scaffolds.
• Human renal epithelial cells were effectively repopulated within the scaffolds.
• Cell growth and viability were monitored and assessed through various assays.
• The study demonstrates the potential for 3D cell culture techniques in organ regeneration.

Conclusions

• The protocol provides a framework for kidney tissue engineering.
• Repopulation of acellular scaffolds with human cells is feasible.
• Further research is needed to optimize conditions for cell growth and differentiation.

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