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Epithelial Cell Repopulation and Preparation of Rodent Extracellular Matrix Scaffolds for Renal Tissue Development
Published on: August 10, 2015
A noninvasive transfer system for polarized renal tubule epithelial cell sheets using temperature-responsive culture
1Institute of Advanced Biomedical Engineering and Science, Tokyo Women's Medical University, 8-1 Kawada-cho, Shinjuku-ku, Tokyo 162-8666, Japan. tokano@abmes.twmu.ac.jp
European Cells & Materials
|August 10, 2005
Summary
Researchers developed a novel cell sheet transfer method using temperature-responsive polymers for tissue engineering. This technique noninvasively moves intact cell sheets while preserving crucial cell polarity and functions.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Cell Biology
Background:
- Maintaining cell polarity is crucial for epithelial tissue function.
- Traditional methods like trypsinization can disrupt cell polarity and structure.
- Tissue engineering requires methods for seamless cell sheet transplantation.
Purpose of the Study:
- To develop a noninvasive method for transferring intact cell sheets.
- To evaluate the preservation of cell polarity and function after transfer.
- To assess the utility of this method in tissue engineering applications.
Main Methods:
- Utilized temperature-responsive culture dishes with poly(N-isopropylacrylamide).
- Harvested confluent cell sheets via temperature reduction.
- Transferred cell sheets to various substrates and analyzed polarity using immunohistochemistry and electron microscopy.
- Confirmed tight junction integrity via occludin immunoblotting.
Main Results:
- Successfully transferred intact cell sheets of Madin-Darby canine kidney cells and human renal proximal tubule epithelial cells.
- Preserved apical-basal cell polarity, including the distribution of key transporters (Na+/K+-ATPase, GLUT-1, SGLT-1, aquaporin-1) and enzymes.
- Observed formation of microvilli and tight junctions in transferred cell sheets.
- Confirmed occludin integrity, indicating functional tight junctions.
Conclusions:
- The temperature-responsive cell sheet transfer method effectively preserves epithelial cell polarity and function.
- This noninvasive technique is suitable for tissue engineering and studying epithelial cell biology.
- The method offers a promising alternative to enzymatic dissociation for cell sheet manipulation.
