Novel extracellular matrix for cell sheet recovery using genetically engineered elastin-like protein.
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 degrees 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.
Mie
Masayasu
M
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 4259 Nagatsuta, Midori-ku, Yokohama 226-8501, Japan.
Mizushima
Yasunori
Y
Kobatake
Eiry
E
D001483:Base Sequence; D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D003470:Culture Media / Q000737:chemistry; D004549:Elastin / Q000737:chemistry*; D005109:Extracellular Matrix / Q000378:metabolism*; D005818:Genetic Engineering / Q000379:methods*; D006801:Humans; D008969:Molecular Sequence Data; D009842:Oligopeptides / Q000737:chemistry*; D010957:Plasmids / Q000378:metabolism; D015202:Protein Engineering / Q000379:methods*; D011506:Proteins / Q000737:chemistry; D013696:Temperature
D001672:Biocompatible Materials; D003470:Culture Media; D009842:Oligopeptides; D011506:Proteins; C047981:arginyl-glycyl-aspartic acid; D004549:Elastin