title: Individual evaluation of cardiac marker expression and self-beating during cardiac differentiation of P19CL6 cells on different culture substrates.
abstract: Cell-based therapies using self-beating cardiomyocytes have been attracting great attention for use in cardiac regeneration, although an effective procedure to improve cardiac differentiation and self-beating induction is required. The purpose of this study is to clarify the effect of the culture substrate on cardiac maturation by separately evaluating the cardiac differentiation step and the beating induction step in vitro. To this end, the well-studied cardiomyocyte-like progenitor cell line P19CL6 and neonatal cardiomyocytes (NCMs) were selected and cultured on substrates coated with collagen type I (Col-I), gelatin (Gel), fibronectin (FN), or poly-l-lysine (PLL). It was found that the cardiac differentiation step, which was assessed using cardiac marker gene expression (GATA-binding protein 4 (GATA4), myocyte-specific enhancer factor 2D (MEF2D), and hyperpolarization-activated cyclic nucleotide-gated potassium channel 4 (HCN4)) in the P19CL6 embryonal carcinoma cells, was greatly enhanced on Col-I, Gel, and PLL. In contrast, the spontaneous beating step, which was directly assessed by counting the beating colonies and measuring contractile protein gene expression (α-myosin heavy chain (α-MHC), troponin C type 1 (TnC1), and troponin T type 2 (TnT2)) in the rat NCMs, was enhanced on the FN and PLL surfaces. In the present study, for the first time, it was found that PLL enhances both the cardiac differentiation and the beating induction steps of cardiac maturation, which can aid in preparing beating cardiomyocytes for regenerative medicine. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1166-1174, 2017.
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.|JST-CREST, 5 Sanbancho, Chiyoda-ku, Tokyo, 102-0075, Japan.
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamaoka
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.|JST-CREST, 5 Sanbancho, Chiyoda-ku, Tokyo, 102-0075, Japan.
authors.forename: Mitsuhi
authors.initials: M
authors.lastname: Hirata
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.|Department of Materials Chemistry, Ryukoku University, 1-5 Seta, Otsu, Shiga, 520-2194, Japan.
authors.forename: Takaaki
authors.initials: T
authors.lastname: Dan
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.|JST-CREST, 5 Sanbancho, Chiyoda-ku, Tokyo, 102-0075, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Yamashita
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.
authors.forename: Akihisa
authors.initials: A
authors.lastname: Otaka
authors.affiliation: Department of Materials Chemistry, Ryukoku University, 1-5 Seta, Otsu, Shiga, 520-2194, Japan.
authors.forename: Takahiko
authors.initials: T
authors.lastname: Nakaoki
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.
authors.forename: Azizi
authors.initials: A
authors.lastname: Miskon
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.|JST-CREST, 5 Sanbancho, Chiyoda-ku, Tokyo, 102-0075, Japan.
authors.forename: Sachiro
authors.initials: S
authors.lastname: Kakinoki
authors.affiliation: Department of Biomedical Engineering, National Cerebral and Cardiovascular Center Research Institute, 5-7-1 Fujishirodai, Suita, Osaka, 565-8565, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Mahara
mesh_terms: D000818:Animals; D000943:Antigens, Differentiation / Q000096:biosynthesis*; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D032386:Myoblasts, Cardiac / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D051381:Rats
chemical_list: D000943:Antigens, Differentiation
keywords: P19CL6; cardiac differentiation; neonatal cardiomyocyte; poly-l-lysine; self-beating