title: Development of a cell culture surface conversion technique using alginate thin film for evaluating effect upon cellular differentiation.
abstract: Here, we sought to develop a cell culture surface conversion technique that would not damage living cells. An alginate thin film, formed on a glass plate by spin coating of sodium alginate solution and dipping into calcium chloride solution, was used to inhibit adhesion of cells. The film could be removed by ethylenediaminetetraacetate (EDTA) at any time during cell culture, permitting observation of cellular responses to conversion of the culture surface in real time. Additionally, we demonstrated the validity of the alginate thin film coating method and the performance of the film. The thickness of the alginate thin film was controlled by varying the rotation speed during spin coating. Moreover, the alginate thin film completely inhibited the adhesion of cultured cells to the culture surface, irrespective of the thickness of the film. When the alginate thin film was removed from the culture surface by EDTA, the cultured cells adhered to the culture surface, and their morphology changed. Finally, we achieved effective differentiation of C2C12 myoblasts into myotube cells by cell culture on the convertible culture surface, demonstrating the utility of our novel technique.
authors.affiliation: Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 096-8555, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Nakashima
authors.affiliation: Graduate School of Science and Engineering, Yamaguchi University, 2-16-1 Tokiwadai, Ube 755-8611, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Tsusu
authors.affiliation: Graduate School of Science and Engineering, Yamaguchi University, 2-16-1 Tokiwadai, Ube 755-8611, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Minami
authors.affiliation: Graduate School of Science and Technology, Kumamoto University, 2-39-1 Kurokami, Chuo-ku, Kumamoto 096-8555, Japan.
authors.forename: Y
authors.initials: Y
authors.lastname: Nakanishi
mesh_terms: D000464:Alginates / Q000737:chemistry*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation; D002460:Cell Line; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D008567:Membranes, Artificial; D051379:Mice; D032446:Myoblasts / Q000166:cytology / Q000378:metabolism*; D013499:Surface Properties
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D008567:Membranes, Artificial; D020723:Glucuronic Acid