In vitro production of insulin-responsive skeletal muscle tissue from mouse embryonic stem cells by spermine-induced differentiation method

• Published in: Human cell : official journal of Human Cell Research Society Vol. 30; no. 3; pp. 162 - 168
• Main Authors: Saito, Mikako, Ishida, Ayano, Nakagawa, Shota
• Format: Journal Article
• Language: English
• Published: Tokyo Springer Japan 01.07.2017; Springer Nature B.V
• Subjects: Animals; Biomedical and Life Sciences; Cell Biology; Cell Differentiation - drug effects; Cells, Cultured; Confocal microscopy; Embryo cells; Embryoid Bodies - cytology; Embryonic Stem Cells - cytology; Glucose; Glucose - metabolism; Gynecology; Insulin; Insulin - pharmacology; Life Sciences; Mice; Muscle, Skeletal - cytology; Muscle, Skeletal - metabolism; Musculoskeletal system; Oncology; Rapid Communication; Reproductive Medicine; Rodents; Skeletal muscle; Skeleton; Spermine; Spermine - pharmacology; Stem cell transplantation; Stem Cells; Surgery; Insulin-stimulated glucose uptake; Skeletal muscle tissue; Multilayer myotube sheet; Spermine; Fluorescent glucose analog
• ISSN: 1749-0774; 0914-7470; 1749-0774
• DOI: 10.1007/s13577-017-0176-8

The treatment of an embryoid body with spermine for a short duration can trigger the generation of a 3-dimensional multilayer myotube sheet (MMTS) that shows pulsatile activity. MMTS was previously characterized as a model of skeletal muscle tissue. In the present work, the insulin responsiveness of MMTS was investigated because it is an essential function for a model of skeletal muscle. The glucose uptake activity of MMTS was analyzed by confocal microscopy using fluorescent glucose analogs, namely 2-[ N -(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]-2-deoxy- d -glucose (2-NBDG) and its l -glucose counterpart, 2-NBDLG. The specific uptake rate of glucose was estimated from the difference between the fluorescent signals of 2-NBDG and 2-NBDLG. It was enhanced by insulin stimulation to 3.6 times higher than the control without insulin, and this insulin responsiveness was maintained for 5 days. The advantages of the 3-dimensional structure of MMTS are discussed in the contexts of its potential in vivo and in vitro uses.