title: 8-Methyltryptanthrin-induced differentiation of P19CL6 embryonal carcinoma cells into spontaneously beating cardiomyocyte-like cells.
abstract: Enhancement of cardiac differentiation is critical to stem cell transplantation therapy for severe ischemic heart disease. The aim of this study was to investigate whether several derivatives of tryptanthrin (1), extracted from the medicinal plant Polygonum tinctorium, induce the differentiation of P19CL6 mouse embryonal carcinoma cells into beating cardiomyocyte-like cells. P19CL6 cells were cultured in α-MEM supplemented with 10% FBS including a test compound or vehicle. Drug-induced differentiation was assessed by measuring the number of beating and nonbeating aggregates and the area of beating aggregates, and the expression of genes involved in cardiac differentiation was evaluated by real-time PCR. A 1 μM concentration of 8-methyltryptanthrin (2) induced the differentiation of P19CL6 cells into cardiomyocyte-like cells to a significantly greater degree than 1% dimethyl sulfoxide (DMSO), a conventional differentiation inducer of P19CL6 cells. Furthermore, 2 strongly increased both the number and the area of spontaneously beating aggregates in comparison with DMSO. Two distinct genes of the calcium channel family, Cav1.2 and Cav3.1, underlying cardiac automaticity were significantly expressed in the presence of 2. Gap junction genes GJA1 and GJA5 contributing to the synchronized contraction of the myocardium were also induced significantly by 2. These results suggest that 2 successfully differentiated P19CL6 cells into spontaneously beating cardiomyocyte-like cells by activating the gene expression of pacemaker channels and gap junctions.
authors.affiliation: Department of Pharmacology, Hirosaki University Graduate School of Medicine , 5 Zaifu-cho, Hirosaki 036-8562, Japan.
authors.forename: Kazuhiko
authors.initials: K
authors.lastname: Seya
authors.forename: Asako
authors.initials: A
authors.lastname: Yamaya
authors.forename: Sayuri
authors.initials: S
authors.lastname: Kamachi
authors.forename: Mana
authors.initials: M
authors.lastname: Murakami
authors.forename: Haruo
authors.initials: H
authors.lastname: Kitahara
authors.forename: Jun
authors.initials: J
authors.lastname: Kawakami
authors.forename: Ken
authors.initials: K
authors.lastname: Okumura
authors.forename: Manabu
authors.initials: M
authors.lastname: Murakami
authors.forename: Shigeru
authors.initials: S
authors.lastname: Motomura
authors.forename: Ken-Ichi
authors.initials: K
authors.lastname: Furukawa
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D054278:Embryonal Carcinoma Stem Cells; D051379:Mice; D015394:Molecular Structure; D009206:Myocardium / Q000166:cytology; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects*; D009930:Organic Chemicals; D010946:Plants, Medicinal / Q000737:chemistry*; D028502:Polygonum / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011799:Quinazolines / Q000737:chemistry / Q000302:isolation & purification / Q000494:pharmacology*
chemical_list: C000591082:8-methyltryptanthrin; D009930:Organic Chemicals; D011799:Quinazolines; C420642:alpha minimal essential medium; D004121:Dimethyl Sulfoxide