title: A lower volume culture method for obtaining a larger yield of neuron-like cells from mesenchymal stem cells.
abstract: Mesenchymal stem cells (MSCs) represent a promising cell source for stem cell therapy to replace neurons damaged by neurodegenerative diseases. A system designed for in vitro neuronal differentiation of MSCs is an indispensable technique, which provides MSC-derived functional neurons for cell-replacement therapies and valuable information in pre-clinical research. This study investigated the effects of reducing the volume of neural induction medium on cell viability and neural differentiation of MSCs. When MSCs were differentiated in low volumes of neural induction medium, rather than using the conventional method, the cell density on culture dishes significantly increased. The % cell death, including apoptosis and necrosis, was significantly lower in the lower volume method than in the conventional method. There were no significant differences between the lower volume and conventional methods in the expression levels of the neuronal marker genes. In an analysis of immunostaining for a mature neuronal marker, no significant difference was detected between the media volumes. These findings demonstrate that neuronal induction of MSCs in low volumes of differentiation medium promoted survival during differentiation and resulted in larger numbers of MSC-derived neurons, compared to the conventional method. This novel lower volume method offers both financial and cell-yield advantages over the conventional method.
authors.affiliation: Department of Communication Disorders, Health Sciences University of Hokkaido School of Psychological Science, 2-5 Ainosato, Kita-ku, Sapporo, Hokkaido, 002-8072, Japan. shimo@hoku-iryo-u.ac.jp.|Department of Communication Disorders, Health Sciences University of Hokkaido School of Rehabilitation Sciences, 1757 Kanazawa, Tobetsu, Hokkaido, 061-0293, Japan. shimo@hoku-iryo-u.ac.jp.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Shimomura
authors.affiliation: Department of Anatomy and Cell Biology, Gunma University School of Medicine, 3-39-22 Showa-machi, Maebashi, Gunma, 371-8511, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Iizuka-Kogo
authors.affiliation: Laboratory of Molecular Biology and Histochemistry, Fujita Health University Joint Research Laboratory, 1-98 Kutsukake-cho, Toyoake, Aichi, 470-1192, Japan.
authors.forename: Naoki
authors.initials: N
authors.lastname: Yamamoto
authors.affiliation: Department of Anatomy I, Fujita Health University School of Medicine, 1-98 Kutsukake-cho, Toyoake, Aichi, 470-1192, Japan.
authors.forename: Ryuji
authors.initials: R
authors.lastname: Nomura
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects; D008810:Mice, Inbred C57BL; D009474:Neurons / Q000166:cytology* / Q000187:drug effects
chemical_list: D003470:Culture Media
keywords: A lower volume culture method; In vitro neuronal differentiation; Mesenchymal stem cell; Neural differentiation