SHIMIZU Tatsuya    Department   Graduate School of Medical Science, Graduate School of Medical Science    Position   Professor
Article types	Original article
Language	English
Peer review	Peer reviewed
Title	Terminus-Selective Covalent Immobilization of Heparin on a Thermoresponsive Surface Using Click Chemistry for Efficient Binding of Basic Fibroblast Growth Factor.
Journal	Formal name：Macromolecular bioscience Abbreviation：Macromol Biosci ISSN code：16165195/16165187
Domestic / Foregin	Foregin
Volume, Issue, Page	24(2),pp.e2300307
Author and coauthor	ONODERA Yu†, KOBAYASHI Jun*, MITANI Seiji, HOSODA Chihiro, BANNO Kimihiko, HORIE Kyoji, OKANO Teruo, SHIMIZU Tatsuya, SHIMA Midori, TATSUMI Kohei*
Publication date	2024/02
Summary	Cell therapy using endothelial cells (ECs) has great potential for the treatment of congenital disorders, such as hemophilia A. Cell sheet technology utilizing a thermoresponsive culture dish is a promising approach to efficiently transplant donor cells. In this study, a new method to prepare terminus-selective heparin-immobilized thermoresponsive culture surfaces is developed to facilitate the preparation of EC sheets. Alkynes are introduced to the reducing terminus of heparin via reductive amination. Cu-catalyzed azide-alkyne cycloaddition (CuAAC) facilitates efficient immobilization of the terminus of heparin on a thermoresponsive surface, resulting in a higher amount of immobilized heparin while preserving its function. Heparin-immobilized thermoresponsive surfaces prepared using CuAAC exhibit good adhesion to human endothelial colony-forming cells (ECFCs). In addition, upon further binding to basic fibroblast growth factor (bFGF) on heparin-immobilized surfaces, increased proliferation of ECFCs on the surface is observed. The confluent ECFC monolayer cultured on bFGF-bound heparin-immobilized thermoresponsive surfaces exhibits relatively high fibronectin accumulation and cell number and detaches at 22 °C while maintaining the sheet-like structure. Because heparin has an affinity for several types of bioactive molecules, the proposed method can be applied to facilitate efficient cultures and sheet formations of various cell types.
DOI	10.1002/mabi.202300307
PMID	37774391