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Novel extracellular matrix for cell sheet recovery using genetically engineered elastin‐like protein
Author(s) -
Mie Masayasu,
Mizushima Yasunori,
Kobatake Eiry
Publication year - 2008
Publication title -
journal of biomedical materials research part b: applied biomaterials
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.665
H-Index - 108
eISSN - 1552-4981
pISSN - 1552-4973
DOI - 10.1002/jbm.b.31019
Subject(s) - pentapeptide repeat , extracellular matrix , elastin , materials science , sequence (biology) , genetically engineered , tissue engineering , cell , biophysics , extracellular , microbiology and biotechnology , matrix (chemical analysis) , biology , peptide , biochemistry , composite material , genetics , gene
Elastin‐like peptides (ELPs) sequences are repeats of the pentapeptide GVGVP, and they have the ability to coaggregate reversibly, depending on the temperature. By exploiting this characteristic, a novel extracellular matrix protein (ECM) containing ELP was developed genetically to harvest a cell sheet from a culture dish. One of the ELP constructs, G288, consisted of 288 repeats of the sequence GVGVGP (G); it was attached to a hydrophobic dish surface. Next, cells with the sequence His‐G36RG36, which has a His tag and an RGD sequence (R) that promotes attachment of the cell between the G36 sequences, consisted of 36 repeats of the sequence GVGVP, were added to the dish. After these cells became confluent, the temperature was changed to 20°C in order to reverse the coaggregation. At this temperature, cells could be detached from the dish as a cell sheet. This genetically engineering method for construction of thermoresponsive ECM would be suitable to modify ECM with further functional domains. © 2007 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 2008