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Tissue Engineering: Construction of a Multicellular 3D Scaffold for the Delivery of Layered Cell Sheets
Published on: October 3, 2014
Cryopreservation of tissue-engineered epithelial sheets in trehalose
Fanfan Chen1, Wenjie Zhang, Wei Wu
1Department of Plastic and Reconstructive Surgery, Shanghai 9th People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai Key Laboratory of Tissue Engineering, 639 Zhi Zao Ju Road, Shanghai 200011, PR China.
Biomaterials
|September 6, 2011
Summary
Trehalose improves cryopreservation of tissue-engineered skin. This method enhances keratinocyte viability and wound healing properties, offering a superior alternative to dimethyl-sulfoxide (DMSO) for preserving epidermal membranes.
Area of Science:
- Biomaterials Science
- Regenerative Medicine
- Tissue Engineering
Background:
- Optimized storage methods are crucial for clinical applications of tissue-engineered epidermal membranes.
- Dimethyl-sulfoxide (DMSO) is commonly used for cryopreserving skin grafts and artificial skin substitutes.
- Evaluating alternative cryoprotectants is essential for improving the efficacy of tissue-engineered products.
Purpose of the Study:
- To investigate the efficacy of extracellular trehalose as a cryoprotectant for human tissue-engineered epidermal membranes.
- To compare trehalose-based cryopreservation with conventional DMSO methods.
- To optimize trehalose concentration for cryopreservation of artificial epidermal substitutes.
Main Methods:
- Keratinocyte (KC) viability, proliferation, and marker expression were assessed after cryopreservation with trehalose and DMSO.
- Trehalose concentration was optimized based on cellular activity post-cryopreservation.
- Artificial epidermal substitutes were cryopreserved using optimized trehalose and transplanted into nude mice to evaluate wound healing properties.
Main Results:
- Trehalose cryopreservation enhanced human KC viability in suspension and artificial skin substitutes.
- Optimized trehalose concentration (0.4m) was determined.
- Trehalose cryopreservation led to faster recovery of EGF and TGF-β1 secretion by KCs post-thawing.
- Trehalose-cryopreserved artificial skin demonstrated comparable wound repair to non-cryopreserved controls in nude mice.
- Significantly enhanced engraftment and wound closure were observed with trehalose compared to DMSO cryopreservation.
Conclusions:
- Extracellular trehalose is an effective cryoprotectant for tissue-engineered epidermal membranes.
- Trehalose improves cryopreservation outcomes, including cell viability and wound healing capacity.
- Trehalose offers a promising alternative to DMSO for the clinical storage of engineered skin tissues.
