Cardiac differentiation of embryonic stem cells by substrate immobilization of insulin-like growth factor binding protein 4 with elastin-like polypeptides.
The establishment of cardiomyocyte differentiation of embryonic stem cells (ESCs) is a useful strategy for cardiovascular regenerative medicine. Here, we report a strategy for cardiomyocyte differentiation of ESCs using substrate immobilization of insulin-like growth factor binding protein 4 (IGFBP4) with elastin-like polypeptides. Recently, IGFBP4 was reported to promote cardiomyocyte differentiation of ESCs through inhibition of the Wnt/β-catenin signaling. However, high amounts of IGFBP4 (approximately 1 μg/mL) were required to inhibit the Wnt/β-catenin signaling and induce differentiation to cardiomyocytes. We report herein induction of cardiomyocyte differentiation using IGFBP4-immobilized substrates. IGFBP4-immobilized substrates were created by fusion with elastin-like polypeptides. IGFBP4 was stably immobilized to polystyrene dishes through fusion of elastin-like polypeptides. Cardiomyocyte differentiation of ESCs was effectively promoted by strong and continuous inhibition of Wnt/β-catenin signaling with IGFBP4-immobilized substrates. These results demonstrated that IGFBP4 could be immobilized using fusion of elastin-like polypeptides. Our results also demonstrate that substrate immobilization of IGFBP4 is a powerful tool for differentiation of ESCs into cardiomyocytes. These findings suggest that substrate immobilization of soluble factors is a useful technique for differentiation of ESCs in regenerative medicine and tissue engineering.
Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-57, 4259 Nagatsuta-cho, Midori-ku, Yokohama 226-8501, Japan.
Ayaka
A
Minato
Hirohiko
H
Ise
Mitsuaki
M
Goto
Toshihiro
T
Akaike
D000818:Animals; D001704:Biopolymers / Q000378:metabolism; D015153:Blotting, Western; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D004549:Elastin / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000166:cytology*; D055767:Immobilized Proteins / Q000737:chemistry*; D047468:Immunoprecipitation; D018974:Insulin-Like Growth Factor Binding Protein 4 / Q000737:chemistry*; D051379:Mice; D008809:Mice, Inbred C3H; D032383:Myocytes, Cardiac / Q000166:cytology*; D010455:Peptides / Q000378:metabolism; D010957:Plasmids; D011994:Recombinant Proteins; D015398:Signal Transduction; D023822:Tissue Engineering; D051153:Wnt Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D051176:beta Catenin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism
D001704:Biopolymers; D055767:Immobilized Proteins; D018974:Insulin-Like Growth Factor Binding Protein 4; D010455:Peptides; D011994:Recombinant Proteins; D051153:Wnt Proteins; D051176:beta Catenin; C502906:poly(glycyl-valyl-glycyl-valyl-prolyl); D004549:Elastin