title: A non-destructive culturing and cell sorting method for cardiomyocytes and neurons using a double alginate layer.
abstract: A non-destructive method of collecting cultured cells after identifying their in situ functional characteristics is proposed. In this method, cells are cultivated on an alginate layer in a culture dish and released by spot application of a calcium chelate buffer that locally melts the alginate layer and enables the collection of cultured cells at the single-cell level. Primary hippocampal neurons, beating human embryonic stem (hES) cell-derived cardiomyocytes, and beating hES cell-derived cardiomyocyte clusters cultivated on an alginate layer were successfully released and collected with a micropipette. The collected cells were recultured while maintaining their physiological function, including beating, and elongated neurites. These results suggest that the proposed method may eventually facilitate the transplantation of ES- or iPS-derived cardiomyocytes and neurons differentiated in culture.
authors.affiliation: On-chip Cellomics Project, Kanagawa Academy of Science and Technology, Kawasaki, Kanagawa, Japan.
authors.forename: Hideyuki
authors.initials: H
authors.lastname: Terazono
authors.forename: Hyonchol
authors.initials: H
authors.lastname: Kim
authors.forename: Masahito
authors.initials: M
authors.lastname: Hayashi
authors.forename: Akihiro
authors.initials: A
authors.lastname: Hattori
authors.forename: Fumimasa
authors.initials: F
authors.lastname: Nomura
authors.forename: Tomoyuki
authors.initials: T
authors.lastname: Kaneko
authors.forename: Kenji
authors.initials: K
authors.lastname: Yasuda
mesh_terms: D000464:Alginates / Q000737:chemistry*; D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000166:cytology*; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006624:Hippocampus / Q000166:cytology*; D006801:Humans; D032383:Myocytes, Cardiac / Q000166:cytology*; D016501:Neurites; D051381:Rats; D017208:Rats, Wistar
chemical_list: D000464:Alginates; D006603:Hexuronic Acids; D020723:Glucuronic Acid