title: Br-DIF-1 accelerates dimethyl sulphoxide-induced differentiation of P19CL6 embryonic carcinoma cells into cardiomyocytes.
abstract: OBJECTIVE
  Stem cell transplantation therapy is a promising option for treatment of severe ischaemic heart disease. Dimethyl sulphoxide (DMSO) differentiates P19CL6 embryonic carcinoma cells into cardiomyocyte-like cells, but with low differentiation capacity. To improve the degree of this differentiation, we have assessed several derivatives of the differentiation-inducing factor-1 (DIF-1), originally found in the cellular slime mould Dictyostelium discoideum, on P19CL6 cells.
  
  
  METHODS
  P19CL6 cells were cultured with each derivative and 1% DMSO for up to 16 days. Differentiation was assessed by measuring the number of beating and non-beating aggregates, and the expression of genes relevant to cardiac tissue. The mechanism of action was investigated using a T-type Ca(2+) channel blocker.
  
  
  RESULTS
  Of all the DIF-1 derivatives tested only Br-DIF-1 showed any effects on cardiomyocyte differentiation. In the presence of 1% DMSO, Br-DIF-1 (0.3-3 µM) significantly and dose-dependently increased the number of spontaneously beating aggregates compared with 1% DMSO alone, by day 16. Expression of mRNA for T-type calcium channels was significantly increased by Br-DIF-1 + 1% DMSO compared with 1% DMSO alone. Mibefradil (a T-type Ca(2+) channel blocker; 100 nM) and a small interfering RNA for the T-type Ca(2+) channel both significantly decreased the beating rate of aggregates induced by Br-DIF-1 + 1% DMSO.
  
  
  CONCLUSIONS
  Br-DIF-1 accelerated the differentiation, induced by 1% DMSO, of P19CL6 cells into spontaneously beating cardiomyocyte-like cells, partly by enhancing the expression of the T-type Ca(2+) channel gene.
authors.affiliation: Department of Pharmacology, Hirosaki University Graduate School of Medicine, Hirosaki, Japan.
authors.forename: K
authors.initials: K
authors.lastname: Seya
authors.forename: K
authors.initials: K
authors.lastname: Kanemaru
authors.forename: M
authors.initials: M
authors.lastname: Matsuki
authors.forename: K
authors.initials: K
authors.lastname: Hongo
authors.forename: H
authors.initials: H
authors.lastname: Kitahara
authors.forename: H
authors.initials: H
authors.lastname: Kikuchi
authors.forename: Y
authors.initials: Y
authors.lastname: Oshima
authors.forename: Y
authors.initials: Y
authors.lastname: Kubohara
authors.forename: K
authors.initials: K
authors.lastname: Okumura
authors.forename: S
authors.initials: S
authors.lastname: Motomura
authors.forename: K-I
authors.initials: KI
authors.lastname: Furukawa
mesh_terms: D000818:Animals; D002121:Calcium Channel Blockers / Q000494:pharmacology; D020747:Calcium Channels, T-Type / Q000502:physiology*; D002454:Cell Differentiation / Q000187:drug effects*; D045744:Cell Line, Tumor; D004121:Dimethyl Sulfoxide; D015870:Gene Expression / Q000187:drug effects*; D006588:Hexanones / Q000494:pharmacology*; D020748:Mibefradil / Q000494:pharmacology; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000502:physiology
chemical_list: C572150:Br-DIF-1; C498197:Cacna1g protein, mouse; D002121:Calcium Channel Blockers; D020747:Calcium Channels, T-Type; D006588:Hexanones; C053433:1-((3,5-dichloro)-2,6-dihydroxy-4-methoxyphenyl)-1-hexanone; D020748:Mibefradil; D004121:Dimethyl Sulfoxide