title: Degree of Suppression of Mouse Myoblast Cell Line C₂C12Differentiation Varies According to Chondroitin Sulfate Subtype.
abstract: Chondroitin sulfate (CS), a type of glycosaminoglycan (GAG), is a factor involved in the suppression of myogenic differentiation. CS comprises two repeating sugars and has different subtypes depending on the position and number of bonded sulfate groups. However, the effect of each subtype on myogenic differentiation remains unclear. In this study, we spiked cultures of C₂C12myoblasts, cells which are capable of undergoing skeletal muscle differentiation, with one of five types of CS (CS-A, -B, -C, -D, or -E) and induced differentiation over a fixed time. After immunostaining of the formed myotubes with an anti-MHC antibody, we counted the number of nuclei in the myotubes and then calculated the fusion index (FI) as a measure of myotube differentiation. The FI values of all the CS-treated groups were lower than the FI value of the control group, especially the group treated with CS-E, which displayed notable suppression of myotube formation. To confirm that the sugar chain in CS-E is important in the suppression of differentiation, chondroitinase ABC (ChABC), which catabolizes CS, was added to the media. The addition of ChABC led to the degradation of CS-E, and neutralized the suppression of myotube formation by CS-E. Collectively, it can be concluded that the degree of suppression of differentiation depends on the subtype of CS and that CS-E strongly suppresses myogenic differentiation. We conclude that the CS sugar chain has inhibitory action against myoblast cell fusion.
authors: Veterinary Anatomy, Department of Veterinary Science, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan. waritak@muses.tottori-u.ac.jp.|Laboratory of Basic Veterinary Science, United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi 753-8515, Japan. waritak@muses.tottori-u.ac.jp.
authors: Katsuhiko
authors: K
authors: Warita
authors: Veterinary Anatomy, Department of Veterinary Science, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan. hsk7030@gmail.com.
authors: Nana
authors: N
authors: Oshima
authors: Department of Regional Environment, Faculty of Regional Sciences, Tottori University, Tottori 680-8551, Japan. ntakeda@rs.tottori-u.ac.jp.
authors: Naoko
authors: N
authors: Takeda-Okuda
authors: Department of Regional Environment, Faculty of Regional Sciences, Tottori University, Tottori 680-8551, Japan. jtamura@rs.tottori-u.ac.jp.
authors: Jun-Ichi
authors: JI
authors: Tamura
authors: Veterinary Anatomy, Department of Veterinary Science, Faculty of Agriculture, Tottori University, Tottori 680-8553, Japan. y-hosa@muses.tottori-u.ac.jp.|Laboratory of Basic Veterinary Science, United Graduate School of Veterinary Science, Yamaguchi University, Yamaguchi 753-8515, Japan. y-hosa@muses.tottori-u.ac.jp.
authors: Yoshinao Z
authors: YZ
authors: Hosaka
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects*; D002459:Cell Fusion; D002460:Cell Line; D019765:Chondroitin ABC Lyase / Q000037:antagonists & inhibitors; D002809:Chondroitin Sulfates / Q000737:chemistry / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D007150:Immunohistochemistry; D051379:Mice; D024510:Muscle Development / Q000187:drug effects; D018485:Muscle Fibers, Skeletal / Q000187:drug effects; D032446:Myoblasts / Q000187:drug effects*
chemical_list: D002809:Chondroitin Sulfates; D019765:Chondroitin ABC Lyase
keywords: cell fusion; chondroitin sulfate E; chondroitinase ABC; fusion index; myogenesis