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Novel extracellular matrix for cell sheet recovery using genetically engineered elastin-like protein
Masayasu Mie1, Yasunori Mizushima, Eiry Kobatake
1Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
Journal of Biomedical Materials Research. Part B, Applied Biomaterials
|December 29, 2007
Summary
Researchers engineered a thermoresponsive extracellular matrix (ECM) using elastin-like peptides (ELPs). This novel biomaterial allows for the temperature-dependent harvesting of intact cell sheets from culture dishes, enabling advanced tissue engineering applications.
Area of Science:
- Biomaterials Science
- Tissue Engineering
- Molecular Biology
Background:
- Elastin-like peptides (ELPs) are known for their temperature-dependent coaggregation properties.
- Conventional cell sheet harvesting methods can be labor-intensive and may damage cells.
- Developing novel extracellular matrix (ECM) mimics is crucial for regenerative medicine.
Purpose of the Study:
- To develop a genetically engineered, thermoresponsive ECM for facile cell sheet harvesting.
- To investigate the utility of ELP-based constructs in creating a cell-adhesive surface.
- To demonstrate the temperature-induced detachment of confluent cell sheets.
Main Methods:
- Genetic engineering of ELP constructs (G288 and His-G36RG36) containing specific peptide repeats.
- Coating hydrophobic culture dishes with ELP constructs to create an artificial ECM.
- Culturing cells expressing His-G36RG36 on the ELP-coated surface.
- Inducing cell sheet detachment by lowering the temperature to 20°C.
Main Results:
- Successful fabrication of a thermoresponsive ECM using ELP sequences.
- Demonstrated reversible coaggregation of ELPs based on temperature changes.
- Achieved detachment of confluent cell sheets at 20°C without compromising cell viability.
- The His-G36RG36 sequence effectively promoted cell attachment.
Conclusions:
- Genetically engineered thermoresponsive ECMs offer a novel method for cell sheet harvesting.
- This approach facilitates the creation of intact cell sheets for tissue engineering.
- The ELP-based system is adaptable for incorporating additional functional domains for further ECM modification.
