Tokyo University of Pharmacy and Life Sciences
Talk Session: SESSION 13: PEPTIDE INSPIRED MATERIALS
Date: Thursday, June 16, 2022
Talk Time: 08:50 am - 09:10 am
Talk Title: Development of Cell Adhesive Peptide-Agarose Matrices for Cell Culture
Motoyoshi Nomizu mainly focuses on Laminin, Peptide, Biochemistry, Cell adhesion and Cell biology. His Laminin study combines topics in areas such as Biophysics, Molecular biology, Integrin and Biological activity. His research investigates the connection between Peptide and topics such as Stereochemistry that intersect with problems in Peptide synthesis.
His work on Peptide sequence, Sepharose and Cyclic peptide as part of general Biochemistry study is frequently linked to Chain, bridging the gap between disciplines. The concepts of his Cell adhesion study are interwoven with issues in Tube formation, Neurite and Cell adhesion molecule. His Cell research integrates issues from Cancer research and in vitro.
Extracellular matrix proteins, such as laminin and collagen, and their active peptides are potential candidates for affording cell adhesiveness to non-cell adhesive materials for three-dimensional, 3D, cell culture. Our group has identified various biologically active peptides from laminins by screening more than 3,000 peptides. These peptides recognize various cell surface receptors and have the potential ability to serve as bio-adhesiveness for 3D cell culture matrices. Previously, we demonstrated that "mixing" of syndecan-binding peptides, such as AG73, RKRLQVQLSIRT, provide cell adhesiveness to agarose gels while integrin-binding peptides, such as A99, AGTFALRGDNPQG, do not.
In this study, to conjugate the laminin peptides to agarose, we synthesized agarose-aldehyde by oxidation of primary alcohols of agarose using TEMPO reagent. Two laminin peptides, AG73 and A99, were synthesized with a CGG sequence at N-terminus and conjugated to agarose-aldehyde via thiazolidine formation between the cysteine residue and the aldehyde.
Two-dimensional cell adhesion assay using fibroblasts showed that AG73- and A99-conjugated agarose gels have potent cell adhesion activity. Cell encapsulation experiments revealed that only the A99-agarose gels promote elongation and proliferation of fibroblasts in a 3D environment.
These results suggest that covalent conjugation of the peptides to agarose is more beneficial to develop a peptide/agarose matrix compared to the previous mixing method, and the peptide-conjugated agarose gels are promising as a matrix for 3D cell culture for anchorage-dependent cells.